Journal of Functional Foods 67 (2020) 103867

Contents lists available at ScienceDirect

Journal of Functional Foods

journal homepage: www.elsevier.com/locate/jff

Bacteria as an alternate biofactory for carotenoid production: A review of its T

applications, opportunities and challenges
⁎
Shristi Rama,b, Madhusree Mitraa,b, Freny Shaha, Sushma Rani Tirkeya,b, Sandhya Mishraa,b,
a
Applied Phycology & Biotechnology Division, CSIR - Central Salt and Marine Chemicals Research Institute, Bhavnagar 364002, India
b
Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India

A R T I C LE I N FO A B S T R A C T

Keywords: Carotenoid has gained a reputation amongst researchers for its robust antioxidant capabilities. This review
Bacterial carotenoid reasons the inevitability of carotenoid for a larger role than just dietary supplements. In an era dominated by
Strain improvement chemical or plant-derived carotenoid, bacterial carotenoid provides a compelling forte to be exploited as a
Genetic engineering promising alternative. Bacteria are amazingly resourceful beings; however, their low carotenoid content makes
Nutraceutical
them undesirable for commercial applications. Here, we have reviewed their applications as a strong prophy-
Multidrug resistance
SWOT analysis
lactic agent in the health sector with a myriad of applications. Additionally, various measures for augmenting
carotenoid yield through new-age technologies, like sequential nutrition starvation, the induction of carotenoid
accumulation in microbial cells by employing several stress factors, and the construction of hyper-carotenoid
producing strains through genetic engineering for creating proficient producers have been offered. Finally,
SWOT (Strengths, Weaknesses, Opportunities and Threats) analysis has been presented to perceive the sig-
nificance of four major components involved in its commercialisation.

1. Introduction
Carotenoids are a wide array of natural biomolecules produced by
plants, algae, yeast, fungi and some bacteria. They range in colour from
red, yellow to orange, and belong to the isoprenoid subfamily. Animals
are unable to synthesise carotenoids and depend exclusively on their
diet for their uptake. However, Arizona University researchers have
reported that aphids are capable of synthesising carotenoids by means
of lateral gene transfer (Moran & Jarvik, 2010). Over 1100 different
carotenoids have been discovered with diverse colours and associated
biological-functional properties from varied sources (Yabuzaki, 2017).
In contrast to primary metabolites, secondary metabolites are not in-
volved in the growth, development and reproduction of an organism.
The tendency to absorb the visible spectrum of light energy
(400–500 nm) enables carotenoid to play a crucial role in photo-
quenching and protection from photodamage. They are primarily found
with chlorophyll as an accessory light-harvesting complex in green
plants. Many organisms tend to accumulate carotenoid under biotic and
abiotic stresses through up-regulation of the carotenoid biosynthetic
pathway as a response to evading degradation. Nowadays, stress
mediated carotenoid accumulation is being harnessed as a successful
strategy to synthesise the desired carotenoid. Previously, carotenoids
were explored for their provitamin A activities, and only very recently
did the scientific community realize that they are endowed with a high
antioxidant potential, which enables them to act against life-threa-
tening diseases such as cancer, age-related macular degeneration, re-
tarded tooth or bone development and impaired rough scaly skin in
humans (Rojas-Garbanzo et al., 2017). The potent antioxidant activity
improves the immune response against infections and plays a distinct
role in ecological functions. These bioactive compounds also evade
problems related to photooxidative damage, which otherwise leads to
an adverse effect on vital cellular machinery such as DNA, lipids and
proteins. Carotenoids have gained in popularity not only as nu-
traceuticals and nutricosmetics, but also as biologically active com-
pounds (Supplementary Fig. 1). To the best of our knowledge, this is the
first review of bacterial carotenoids that discusses its varied applica-
tions in the food and feed industries, as well as a therapeutic agent. In
this review, we intend to provide an abridged knowledge of a decade
long journey highlighting the notable research from scientific litera-
tures published over the past 5 years. This review discusses genetic
engineering methodologies, stress-mediated carotenogenesis, various
challenges associated and a SWOT (Strengths, weaknesses, opportu-
nities and threats) analysis to identify the internal and external factors
in the commercialisation of bacterial carotenoids.

⁎
Corresponding author at: Applied Phycology and Biotechnology Division, CSIR-Central Salt and Marine Chemicals Research Institute, Bhavnagar 364002, Gujarat,
India.
E-mail address: smishra@csmcri.res.in (S. Mishra).

https://doi.org/10.1016/j.jff.2020.103867
Received 5 November 2019; Received in revised form 4 February 2020; Accepted 22 February 2020
1756-4646/ © 2020 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/BY-NC-ND/4.0/).
S. Ram, et al.
1.1. The odyssey of carotenoid research
The need to search for naturally derived bioactive compounds with
the capability of impeding ailments encouraged researchers to explore
various plant sources for discovering an unconventional and sustainable
carotenoid source. This field experienced vigorous growth during the
nineteen-seventies, when public awareness developed both towards
health safety and an inclination towards the consumption of naturally
derived pigments over those that were chemically synthesised. This
boosted interest to find potential alternatives, e.g. microalgae, yeasts,
filamentous fungi and bacteria. Thus, making carotenoids became a
priority for the industrial production of various lucrative sectors in-
cluding food, nutraceutical, pharmaceutical and nutricosmetics
(Numan et al., 2018). Plants and algae have long been studied for their
ability to synthesise carotenoids. Recently, bacterial carotenoids have
come into the mainstream due to their opportunity for higher biomass
cultivation in short periods of time, lack of seasonal restrictions, fast
and easy cultivation, wide variety of colours, nontoxicity, easier ex-
traction process and biodegradable end product with multiple clinical
applications.
2. Chemical structure: the essence of its concomitant activities
Carotenoids are composed of symmetric C40 tetraterpenoid hydro-
carbon compounds consisting of eight isoprene units (polyisoprene),
each having a C5 isoprene member generating a C40 carotenoid com-
pound (Aryee, Agyei, & Akanbi, 2018). The most specific character of
carotenoids is the presence of alternate conjugated bonds. Polyene
structures comprising ≥ 9 conjugated bond systems are known to ex-
hibit colour. Their absorption within the visible range, i.e. 400–500 nm,
is a result of their strong electronic transition from the S0 ─►S2 stage,
which corresponds to an intense colour ranging from yellow, orange
and red (Zigmantas, Hiller, Sundström, & Polívka, 2002). Since the
pigment differs in the number and stereochemistry of the double con-
jugated bonds in the chromophore, there is an increase in the intensity
of the colour from yellow to red along with the increase in the number
of conjugated bonds (Al-Babili & Bouwmeester, 2015). Based on the
presence of oxygen, carotenoids are broadly classified as (i) xanthophyll
(oxygen-bearing) comprising lutein, zeaxanthin, neoxanthin, violax-
anthin and α − and β − cryptoxanthin, and (ii) carotene (composed of
C and H atoms), which lycopene, β-carotene etc belong to. Long-chain
conjugated double bonds hold an electron-rich environment in car-
otenoids, which personifies them to be a proficient antioxidant com-
pound. This, however, renders carotenoids to be highly reactive to-
wards oxygen, heat and light (Britton, Liaaen-Jensen, & Pfander, 1995).
The chemical characteristics of the different carotenoids are shown in
Supplementary Table 1.
3. Degradation and isomerization
Carotenoids, by virtue of their electron-rich polyene chain, are
highly susceptible to degradation upon exposure to light, heat, oxygen,
acids and alkaline bases (Xiao et al., 2018). These factors lead to their
chemical transformations into cis-trans isomerization through the
shifting of bonds and electrons (Supplementary Fig. 2). Upon exposure
to light, oxygen and heat, the carotenoid degradation pathway under-
goes mainly three chemical changes (i) oxidative decomposition (ii)
cis–trans isomerization and (iii) isomerization of 5, 6–epoxides and 5,
8–epoxides. Oxidation is an irreversible process involving a free-radical
reaction resulting in the degradation of products such as epoxide,
apocarotenoid and apocarotenal which is accountable for major car-
otenoid degradation (Pénicaud, Achir, Dhuique-Mayer, Dornier, &
Bohuon, 2011). Further fragmentation yields low molecular molecules
such as methylheptenone, laevulinic aldehyde, probably glyoxal and
acetone for lycopene oxidation, as well as 5,6-epoxy-β-ionone, ionene,
β-cyclocitral, dihydroactinidiolidem β-ionone and 4-oxo-β-ionone for
2
Journal of Functional Foods 67 (2020) 103867
β-carotene oxidation. These losses can be neutralised through the ad-
dition of carbonate, pyridine and dimetilalanine. Stereoisomerism also
plays a key role in the functional activity of carotenoids, which influ-
ences their solubility and absorbance efficiency. Trans forms have a
higher tendency to crystallise and aggregate because they are rigid.
Over time, trans β − carotene forms were rendered more valuable than
their counterparts, cis-β − carotene forms. Ben-Amotz and Levy (1996)
provided the reason behind the higher activity of trans forms with their
capability of becoming absorbed favourably over 9-cis-β − carotene in
humans and ferrets (Ben-Amotz & Levy, 1996). However, in the case of
lycopene (which is not associated with vitamin A activity), cis forms are
more bioavailable than trans-lycopene in ferrets (Rodriguez-Amaya &
Kimura, 2004).
4. The occurrence and biosynthesis of carotenoids
Some of the natural carotenoids and their functional groups are
listed in Supplementary Table 2. Carotenoid biosynthesis occurs via two
different pathways, as discovered earlier and is the well-studied me-
valonate pathway (MVP) in plants (Rodrıguez-Concepción & Boronat,
2002), as well as a newly discovered methylerythritol 4-phosphate
(MEP) pathway for carotenoid synthesis in eubacteria and Escherichia
coli (Rohmer, Knani, Simonin, Sutter, & Sahm, 1993). Two 5C subunits,
isopentenyl diphosphate (IPP) and its isomeric form dimethylallyl di-
phosphate (DMAPP), are the universal precursors for manufacturing
C40 and C50 carotenoids (Fig. 1). The isomerization of IPP into DMAPP
is carried out by IPP/DMAPP isomerase (IDI). Another enzyme, ger-
anylgeranyl diphosphate (GGPP) synthase, catalyses the addition of
three IPP molecules with one DMAPP molecule to yield the immediate
precursor of the carotenoid biosynthesis, a C20 geranylgeranyl dipho-
sphate (GGPP) molecule. Two GGPP molecules undergo tail-to-tail ad-
dition to form the first parent colourless molecule phytoene (C40),
whose formation establishes the committed step in carotenoid bio-
synthesis. Phytoene undergoes four consecutive reactions mediated by
phytoene desaturase, zeta (ζ)-carotene isomerase, zeta (ζ)-carotene
desaturase and carotene isomerase. Phytoene desaturase catalyses the
first desaturation reaction, yielding ζ–carotene, which undergoes two
more desaturation steps, thus forming the first coloured product, ly-
copene, which is linear. Lycopene is considered to be the backbone of
subsequent customisation reactions yielding 1100 different carotenoid
forms. Cyclization at both the ends of lycopene, through the addition of
β-ionone rings by lycopene β cyclase, forms α–carotene and β–car-
otene. The presence of these β-ionone rings bestows β-carotene with its
pro-vitamin A competence. The subsequent hydroxylation of α–car-
otene and β–carotene into lutein and zeaxanthin occurs through β-ring
hydroxylase and α-ring hydroxylase, respectively.
In brief, carotenoid’s main backbone undergoes the following che-
mical reactions to form around 1100 wide types of carotenoids, (i)
cyclization, resulting in seven different end groups (Ψ, β, ε, γ, κ, Φ and
ϰ), (ii) hydrogenation, bond migration, breaking or linking up of hy-
drocarbon chains, and (iii) oxygenation or methylation (Britton, Liaaen-
Jensen, & Pfander, 1995).
5. Carotenoid market drive and commercial relevance
The global carotenoid market was $1.5 billion in 2017 and is an-
ticipated to reach $2.0 billion in 2020 (McWilliams, 2018). Global
market polls (2018–2024) have estimated that the market share of
carotenoids in food and beverages, pharmaceuticals, cosmetics, animal
feed and dietary supplements were 26.1%, 9.2%, 6.5%, 34.8% and
23.5%, respectively. Currently, around 80–90% of the present car-
otenoid supply is fulfilled by chemical synthesis (Saini & Keum, 2019).
Consequently, the market share of natural carotenoids is considerably
less (24%) compared to synthetic carotenoids (76%) due to their high
costs (Market Research Report, 2016). According to the Deinove bio-
technology company, the relatively low market value of synthetic
S. Ram, et al. Journal of Functional Foods 67 (2020) 103867

carotenoids ($250–$2000 kg−1) seems to be one of the main factors

behind their huge market share, whereas natural plant-based car-
otenoids project a market value of $350-$7500 kg−1 (Deinove, n.d). In
2016, the annual market value of β-carotene, lutein, astaxanthin,
zeaxanthin, canthaxanthin, capsanthin, lycopene, Annatto and β-apo-8-
carotenal ranged between $22.5 and $285 million (McWilliams, 2018).
Since plant-derived carotenoids are costly, scientific interest in bac-
terial carotenoid production has escalated in recent years due to their
economic sustainability and cost-effectiveness.

6. Carotenoid production from bacteria: its scope and significance

Naturally pigmented salt lakes and ponds located in arid regions are
the consequence of microscopic halophiles (the Halobacteriaceae ar-
chae family), diatoms and microalgae. The Grand Prismatic Spring in
Yellowstone National Park in the United States; Lake Natron in
Tanzania; Lake Lago di Tovel in Italy; Lake Retba or ‘Lac Rosa’ in
Senegal; and Lake Hillier in Australia are a few of the well-known lakes
or ponds (Supplementary Fig. 3). Carotenoids such as bacterioruberin,
β-carotene and salinixanthin contribute to the red or orange appearance
of hypersaline lakes and ponds (Oren, 2009). They help protect their
delicate cells from the extremely humid desert sunlight. Carotenoid
production from bacterial sources is an attractive alternative for plant-
based carotenoids, as it ensures greater productivity due to their short
life cycle, round-the-year carotenoid production, the probability of
novel carotenoids and the ease of maintaining controlled conditions
during fermentation. Further, converting a low-cost substrate (in-
dustrial or agricultural waste) into a high-value carotenoid reiterates
our waste-to-wealth commitment. Moreover, the prokaryotic nature of
bacteria makes it easier to produce genetic manipulations compared to
eukaryotic algae and plant systems. Some examples of industrially
important pigment-producing microbes are presented in Table 1. Pop-
ular bacterial species reported for carotenoid production are Flavo-
bacterium spp., Agrobacterium spp., Micrococcus spp., Pseudomonas aer-
uginosa, Serratia marcescens, Chromobacterium spp., Rheinheimera spp.
and Arthrobacter spp. Various applications of bacterial carotenoids
(astaxanthin, β-carotene, zeaxanthin, canthaxanthin, lycopene) in the
field of food, feed and as prophylactic agents against multiple diseases,
as listed in Table 2.
Fig. 1. Biosynthesis pathway involved in carotenogenesis as reported in bac-
teria.

Table 1
List of industrially important carotenoid producing microbes.
Microbes Carotenoid Organism Carotenoid content References

−1
Bacteria β-Cryptoxanthin Brevibacterium linens 0.3 mg L culture (Guyomarc’h, Binet, & Dufossé, 2000)
Bacteria Canthaxanthin Bradyrhizobium sp. 1.3 mg g−1 biomass (Lorquin, Molouba, & Dreyfus, 1997)
−1
Bacteria Canthaxanthin Gordonia jacobeae MV-1 13 mg L (Veiga-Crespo, Blasco, Rosa dos Santos, Poza, & Villa, 2005)
Bacteria Canthaxanthin Dietzia natronolimnaea HS-1 7.67 mg L−1 (Gharibzahedi, Razavi, Mousavi, & Moayedi, 2012)
Bacteria Zeaxanthin Flavobacterium sp. 500 mg L−1 (Sajilata, Singhal, & Kamat, 2008)
Bacteria Zeaxanthin Mesoflavibacter zeaxanthinifaciens – (Asker, Beppu, & Ueda, 2007)
Bacteria Zeaxanthin Zeaxanthinibacter enoshimensis – (Asker, Beppu, & Ueda, 2007)
Bacteria Zeaxanthin Rubritalea squalenifaciens – (Shindo & Misawa, 2014)
Bacteria Zeaxanthin Hanstruepera neustonica – (Hameed et al., 2014)
Bacteria Zeaxanthin Synechocystis sp. PCC 6803 0.9 mg L−1 culture (Lagarde, Beuf, & Vermaas, 2000)
Bacteria Zeaxanthin Sphingobacterium multivorum 10.6 mg L−1 (Bhosale, 2004)
Bacteria Astaxanthin-glycoside Sphingomonas sp. PB304 – (Kim, Kim, Lee, & Lee, 2014)
Bacteria Astaxanthin Brevundimonas sp. N-5 0.36 mg g−1 biomass (Asker, Awad, Beppu, & Ueda, 2012)
Bacteria Astaxanthin Brevundimonas sp. M7 (mutant) 1.3 mg g−1 biomass (Asker et al., 2012)
Bacteria Astaxanthin Brevundimonas sp. SD212 7.4 mg g−1 Lu, Bu, and Liu (2017)
Bacteria Astaxanthin Paracoccus bogoriensis 0.4 mg g−1 biomass (Osanjo, Muthike, Tsuma, Okoth, Bulimo, Luuml, Abraham, Dion, Timmis, &
Golyshin, 2009)
Yeast Astaxanthin Phaffia rhodozyma 6.8 mg L−1 (Batghare, Singh, & Moholkar, 2018)
Yeast Astaxanthin Haematococcus pluvialis 36 mg g-1 (Christian, Zhang, Sawdon, & Peng, 2018)
Microalgae Zeaxanthin Microcystis aeruginosa – (Chen, Li, Wong, Ji, & Jiang, 2005)
Microalgae Zeaxanthin Spirulina – (Toyomizu, Sato, Taroda, Kato, & Akiba, 2001)

3
S. Ram, et al. Journal of Functional Foods 67 (2020) 103867

(Eye Disease Case Control Study Group,

(Zhang, Sun, Sun, Chen, & Chen, 2014)
(Dufossé et al., 2005; Virtamo et al.,

(Viuda-Martos et al., 2014)

(Fasano et al., 2014)
References

2014)

1993)
Protects from age related macular degeneration (AMD) and cataracts, accentuate

Fig. 2. Transverse cell membrane orientation of β-carotene and zeaxanthin.

Antioxidants and inhibit lipid oxidation in liposomes, widely used as Food
Antioxidants and widely used as food colorants, Prevent night blindness

7. Production of bacterial carotenoids

Carotenoids are extremely hydrophobic molecules and are restricted

to the integral parts of complex bacterial cytoplasmic and cell wall
membranes (Fig. 2). The orientation and localisation of carotenoids are
reported to reinforce mechanical strength, flexibility, rigidity to the cell
Feed, pharmaceutical and aquaculture industries

membrane and lower susceptibility to lipid peroxidation for upholding

Prevention against Breast and stomach cancer

cell sustainability under extreme environments (Kirti, Amita, Priti,

Mukesh Kumar, & Jyoti, 2014; Lutnaes, Strand, Pétursdóttir, & Liaaen-
Jensen, 2004; Vila, Hornero-Méndez, Azziz, Lareo, & Saravia, 2019).
Novel carotenoids such as deinoxanthin and thermozeaxanthin have
been reported in Deinococcus radiodurans and Thermus thermophiles,
which were found to aid the isolates in combating extreme environ-
mental stresses such as radiation, oxidation, desiccation and high
temperature (Tian & Hua, 2010). Another extremophilic bacteria Rho-
egg yolk color

dococcus sp. B7740 was reported to synthesise an aromatic carotenoid

Applications

isorenieratene which displayed higher stability after gastrointestinal

colorant

tract ingestion than β-carotene and lutein, thus suggesting its great
promise in food applications (Chen, Xie, Yang, Chen, & Sun, 2018).
Many promising carotenoid-producing bacterial strains have been
Staphylococcus aureus, Vibrio psychroerythrus, Flavobacterium sp., Paracoccus

isolated throughout freshwater and marine waters that possess diverse

carotenoid profiles. Around 30 bacterial isolates belonging to the
Arthrobacter, Flavobacterium, Chryseobacterium and Zobellia genera iso-
lated from King George Island, Antarctica were found to produce ten
different carotenoids, out of which zeaxanthin, β-cryptoxanthin and β-
carotene were common carotenoids (Vila et al., 2019). However,
Agrobacterium aurantiacum, Paracoccus carotinifaciens
List of carotenoids produced by different bacterial strains and their applications.

around 37 potential freshwater bacteria as a source of ketocarotenoids

(astaxanthin and adonixanthin) were discovered by Asker and co-
workers, out of which the FrW-Asx-5 isolate exhibited as much
Monascus roseus, Paracoccus carotinifaciens,

as ~ 46% astaxanthin content (Asker, Awad, Beppu, & Ueda, 2018).

Bacterial growth and carotenoid production is known to be gov-
erned by external factors such as culture conditions and environmental
parameters, which directs their biosynthesis pathways. Laboratory-
scale optimisations using the one factor at a time approach for influ-
encing environmental parameters such as pH, temperature, carbon
Rhodospirillum rubrum

source, nitrogen source, salinity etc., are time-consuming and often

Sphingomonas sp.
Microorganism

produce inconsistent data in large-scale cultivation. Fermenters are thus

xanthinifaciens

a viable alternative to overcome these limitations and have been de-

vised to enhance the carotenoid process development for industrial
production. Fermenters provide a controlled regulation of the en-
vironmental parameters (optimum conditions), which leads to a sig-
Reddish-orange
Yellow- orange

nificant increase in carotenoid production. A feed-batch fermentation

Golden Yellow

study using a recombinant strain of E. coli resulted in a lycopene ac-

cumulation of 220 mg L−1 from a 27 g dry cell weight L−1 (Alper,
Color

Miyaoku, & Stephanopoulos, 2006). Along with enhancing the bacterial

Red

Red

carotenoid production under optimum conditions, using cheap natural

Canthaxanthin

substrates (such as a carbohydrate source) during the fermentation

Astaxanthin
β- carotene

Zeaxanthin
Carotenoid

process could also reduce the overall cost. For instance, an escalated
Lycopene

amount of carotenoid production occurred through the co-cultivation of

Table 2

Rhodutorula rubra and Lactobacillus casei using whey filtrate (Simova,

4
S. Ram, et al. Journal of Functional Foods 67 (2020) 103867

Frengova, & Beshkova, 2003). A successful biotransformation of waste

(Chattopadhyay, Jagannadham, Vairamani, & Shivaji, 1997)

glycerol from the biodiesel industry into a colourless carotenoid (phy-

(Antognoni, Mandrioli, Potente, Saa, & Gianotti, 2019)

toene) and β-carotene was demonstrated by several halophiles isolated
from Lake Letea (Neagu et al., 2019).

(Patthawaro, Lomthaisong, & Saejung, 2019)

8. Role of stressors in carotenogenesis

Fontes, Ruiz-Vázquez, and Murillo (1993)

(Fong, Burgess, Barrow, & Glenn, 2001)

(D’Souza, Altekar, & D’Souza, 1997)

Diverse environmental factors and nutrient composition affect the
regulation of the carotenoid pathway. A stressful environment often
(Zhou, Zhang, & Zhang, 2014)
(Montero-Lobato et al., 2018)

leads to the accumulation of carotenoids as a means of a cellular de-

(Fang, Ku, Lee, & Su, 2010)

fensive response to the prevalent nutrient stress condition (Paliwal

et al., 2017; Ram, Paliwal, & Mishra, 2019). Variations in temperature,
(Wang et al., 2017)

salinity, light and pH, which have been reported to trigger car-
otenogenesis and its biosynthetic pathways in some bacterial species,
Reference

are listed in Table 3. Using glycerol as a cheap carbon source in re-

combinant E. coli has resulted in a 3.3-fold increase in torulene, a potent
anti-cancerous carotenoid (Kot, Błażejak, Gientka, Kieliszek, & Bryś,
2018; Lee, Mijts, & Schmidt-Dannert, 2004), whereas glucose supple-
mentation has repressed torulene yield. In another study, Rhodococcus
Bisanhydrobacterioruberin, monoanhydrobacterioruberin, bacterioruberin production

opacus PD630 accumulated 7.5-fold higher carotenoid concentration

when glycerol was utilized as carbon source (Suwaleerat,
Thanapimmetha, Srisaiyoot, Chisti, & Srinophakun, 2018). Tran-
scriptomics studies in Monascus purpureus YY-1 have confirmed that
carbon starvation stress inhibits the central carbon metabolism, thereby
assisting in up-regulating the carotenoid biosynthesis pathway (Yang
et al., 2015). The impact of stressors, such as variations in carbon, ni-
trogen, phosphorus and oxygen content on other counterparts, are also
well-known for stimulating carotenogenesis. Studies on Neurospora
crassa and Phaffia rhodozyma have shown that exposure to oxygen has
stimulated carotenogenesis, probably through gene regulation (Iigusa,
Increased carotenoid and bacteriochlorophyll

Yoshida, & Hasunuma, 2005; Zhang et al., 2019). The effect of nutrient
limitation and salinity stress resulted in a 3.5-fold accumulation of β-
carotene in Synechocystis sp. 2501 (Paliwal et al., 2015). Another study
4.4 fold increase in total carotenoid

showed improved astaxanthin production when Phaffia rhodozyma was

grown at low ammonium or phosphate levels (Flores-Cotera, Martin, &
Sanchez, 2001). Hence, the prospect of getting a high biomass content
Increased bacterioruberins

Bacterioruberin synthesis

Carotenoid upregulation

and a high carotenoid yield is alluring for evaluating the application of

carC gene regulation

various stress factors for bacterial carotenoid production.

High carotenoid

High carotenoid
High carotenoid
Carotenogenesis

9. Genetic engineering: A boon for strain improvement

The competitive industrial demands limit the use of wild-type

Effect of abiotic stressors on carotenogenesis reported in some bacterial species.

strains for carotenoid production, as they are prone to produce rela-

tively low concentrations of the desired carotenoid compounds, thus
leading to insufficient supplies and making them an unattractive pro-
Rhodopseudomonas faecalis

spect for industrial use due to the associated high costs. Genetic mod-
Rhodopseudomonas sp.

Rhodopseudomonas sp.
Haloferax mediterranei
Haloferax mediterranei

Haloferax mediterranei

ifications are a proven asset in strain improvement in collaboration

Myxococcus xanthus
Micrococcus Roseus

Enterococcus gilvus
Arthrobacter agilis

with mitigating the toxic issues generated by synthetic dye.

Additionally, the knowledge of whole genome sequencing leads to the
identification of biosynthetic pathways which could be altered for the
Sources

focused over-expression of a single desired carotenoid, thereby con-

trolling the concomitant production of unwanted co-products.
Since the bacterial genome is more easily manipulated due to its
simpler genetic design, there is a huge possibility for the development
of a robust and high carotenoid, thus yielding an easily achieved,
Stress condition

30 °C to 5 °C

commercially acclaimed competitive strain. The obstacle, however, lies

25 S m−1
50 g L−1

in the absence of the membrane bound organelles required for the

8000 lx
< 20%

6.8–8.3

functional expression of eukaryotic enzymes (eg. cytocrome P450s).

5 °C
5%

Therefore, to overcome these issues, different strategies have been de-

vised to achieve the metabolic flux for the target molecule; such as
increasing precursor supplies, down-regulating the genes responsible
Oxidative stress

for branched intermediates, thus increasing enzyme specificity, in-

Abiotic stress

Temperature

creasing co-factor levels, substrate channelling and providing an ample

Blue light
Salinity

supply of precursors such as di-methylallyl pyrophosphate (DMAPP)

Table 3

Light

and isopentenyl diphosphate (IPP). High carotenoid productivities can

pH

also be achieved by over-expressing the idi (Isopentenyl-diphosphate

5
S. Ram, et al.
Delta-isomerase) and the dxs (1-deoxy-D-xylulose-5-phosphate syn-
thase) genes in the DXP (1-deoxy-D-xylulose 5-phosphate) pathway
(Choi, Lee, Kim, & Woo, 2010). For example, a maximum lycopene
yield was attained when pyruvate and glyceraldehyde 3-phosphate
(G3P) was over-expressed through up-regulating ppsA (phosphoe-
nolpyruvate/PEP synthase) and pck (PEP carboxykinase) along with the
simultaneous inhibition of pyfFA (Pyk-I and –II) (Farmer & Liao, 2001).
However, further studies on E. coli demonstrates that the down-
regulation of gapA (G3P dehydrogenase) is superior than ppsA over-
expression for lycopene accumulation (Jung et al., 2016). Heterologous
expression enables inserting a carotenoid metabolic route into an ori-
ginally non-carotenoid-forming microorganism, thus turning them into
carotenoid-producing cell-factories with industrial potential. For in-
stance, an E. coli mutant was created by inserting carotenoid biosyn-
thetic genes from Deinococcus wulumuqiensis, which resulted in a 2.2-
fold increase in lycopene production (688 mg L−1), followed by the
wild type (Xu et al., 2018). In another study, inserting a heterologous
mevalonate (MEV) pathway into an originally MEV lacking E. coli strain
spiked with isoprene 20 times more than the wild type (Yang et al.,
2016).
The enhancement of catalytic activity and product specificity is
done using structural-based enzyme engineering strategies for specific
bioconversion efficiency. For example, the promiscuous NphB enzyme
with the ability to prenylate multiple aromatic compounds, was en-
gineered using a protein-docking simulation study. This resulted in the
development of the flux towards geranylated products, with a 3.75-fold
increase (Yang, Park, Park, Eun, & Lee, 2020).
The deletion of sigma factor B (sigB) in Corynebacterium glutamicum,
a yellow-coloured bacteria, favoured the over-expression of sigA factor,
resulting in 2- to 8-fold increase in the decaprenoxanthin and lycopene
yield, respectively, along with the additional synthesis of non-in-
digenous biopigments such as bisanhydrobacteriorberin and β-carotene
(Taniguchi, Henke, Heider, & Wendisch, 2017).
Strategies were employed to alter the genetic sequences using mu-
tagenic techniques such as UV irradiation (7.2 mg L−1) for Micrococcus
roseus PTCC 1411 (Yolmeh, Khomeiri, Ghorbani, Ghaemi, &
Ramezanpour, 2017) and genetic recombination, in silico flux varia-
bility scanning (FVSEOF), the induction of isopropyl-β-D-thiogalacto-
pyranoside (IPTG), and post-segregation killing by the hok/sok system
to create hyper-metabolites and pigment manufacturing strains were
developed and applied to evaluate the carotenoid accumulation in
strains (Yolmeh & Khomeiri, 2016).
Whole-genome sequencing also provides valuable insights into the
genetic make-up of the promising pigment synthesising microorgan-
isms, their biosynthetic pathways and the presence of a myriad of
functional genes, which can help in elucidating the potential of a par-
ticular strain. The identification of genes involved in carotenogenesis
paves the way for elucidating the functional genomics of carotenoid
synthesis. This advancement assists in laying a strong foundation for
further exploratory metabolic engineering activities for strain im-
provement, in turn leading to the production of highly efficient strains
for producing secondary metabolites catering for industrial needs
(Aruldass, Dufossé, & Ahmad, 2018).
The draft genome of the Planococcus maritimus MKU009 strain was
studied to locate and identify carotenoid-producing enzymes such as
phytoene synthase (crtB), phytoene desaturase (crtP), phytoene dehy-
drogenase (crtI) and diapophytoene desaturase (crtN) for ensuring a
thorough understanding of the biosynthesis of the yellow pigment
(Ganapathy, Jayavel, & Natesan, 2016). Well-studied and established
genomes, fermentation techniques, system metabolic engineering tools
and strategies (including genome-scale metabolic models (GEMs) offers
E. coli as a protagonist for most of the genetic manipulation studies for
natural product biosynthesis (Kim, Kim, & Lee, 2017). The metabolic
engineering tools that mediated the orchestration of the carotenoid
biosynthetic pathways in E. coli yielded a 11.95 ± 0.21 mg zeaxanthin
g−1 dry weight (Li, Tian, Shen, & Liu, 2015). The genetically modified
6
Journal of Functional Foods 67 (2020) 103867
E. coli with biosynthetic genes resulted in a higher lycopene content in
strains with an enhanced supply of precursors (Rodríguez-Villalón,
Pérez-Gil, & Rodríguez-Concepción, 2008). It is reported that the en-
gineered strains with overexpressed deoxyxylulose 5-phosphate syn-
thase (DXS) showed an 8-fold increase in lycopene accumulation in the
absence of inducers. High levels of mevalonic acid (MVA + ) operon
enzymes caused interference with the cell metabolism and yielded a 10-
fold lycopene accumulation in E. coli cells. The study further confirmed
the direct regulation of carotenoid production through the
MVA + pathway. A Pseudomonas putida recombination strain was re-
ported to produce up to 239 mg L−1 of zeaxanthin with lecithin addi-
tion during the cell growth (Beuttler et al., 2011). The simultaneous
production of L-lysine (48 g L–1) and astaxanthin (10 mg L−1) was
achieved when Corynebacterium glutamicum was modified through
chromosomal deletion and integration using suicide vector pK19mob-
sacB (Henke, Wiebe, Pérez-García, Peters-Wendisch, & Wendisch,
2018). The phototropic Rhodobacter sphaeroides bacterium was geneti-
cally redesigned to obtain an improved lycopene producing strain with
a final content of 10.32 mg g−1 dry cell weight (88%) (Su et al., 2018).
Carotenoid from the Flavobacterium species is known to consist of
95–99% zeaxanthin. Flavobacterium multivorum is the only bacterial
source that has been commercialised for zeaxanthin production with an
ability to produce 190 mg zeaxanthin L−1 (Shepherd, Dasek, & Carels,
1976).
Enzyme engineering techniques employ the directed evolution of a
target enzyme followed by high-throughput screening using random
mutagenesis in E. coli cells resulting in a 60% increase of isoprene titre
when flux is directed towards 1-deoxy-D-xyloluse-5-phosphate (DXP)
pathway (Lv et al., 2016).
Modulating the gene expression of 1-deoxy-D-xylulose-5-phosphate
synthase (dxs), isopentenyl diphosphate isomerise (idi) and the car-
otenoid biosynthesis gene (crt), gene operon with a screened ribosome
binding site (RBS) in the CAR001 E. coli strain was found to improve the
lycopene content by 32% compared to the parent strain (Sun et al.,
2014). The optimisation of various components such as promoter
strength, RBS strength and 5′-untranslated region sequences are time-
consuming and labour intensive. Thereby, fine-tuning of the gene ex-
pression is of prime importance for a balanced target compound pro-
duction. To achieve this, a novel method has been developed for tuning
the gene expression by generating libraries of tunable intergenic re-
gions (TIGRs) using a high-throughput screening method (Pfleger,
Pitera, Smolke, & Keasling, 2006). TIGR libraries mediate expressions of
the mevalonate (MEV) pathway, yielding a 7-fold (23.16 mg g−1 DCW
zeaxanthin) increase in the MEV production of E. coli (Shen et al.,
2016). Substrate channelling is also an effective strategy that is em-
ployed for the efficient bioconversion of the substrates to the target
molecule. This method transfers the target enzyme to the proximity of
the substrates using protein-protein interactions. It further diminishes
the metabolic burden, the additional by-product formation and the
substrate diffusion. Substrate channelling led the MEV titre to increase
by 77-fold when synthetic protein scaffolds (SH3, PDZ and GBD do-
mains) were hinged around the MEV biosynthetic enzyme (Shen et al.,
2016). Controlling the promoters for balanced upstream and down-
stream process along the DXP pathway resulted in a 4.72-fold increase
in isoprene production (Lv et al., 2016). With the advancement of
computational prediction tools, the use of flux variability scanning
based on the enforced objective flux (FVSEOF) algorithm to identity
amplified gene targets, has enabled E. coli to produce a high content
and a higher productivity of astaxanthin (Park, Binkley, Kim, Lee, &
Lee, 2018).
Apart from bacteria, there are some other microbial sources that are
genetically modified for higher carotenoid accumulation; for example, a
mutant of Rhodotorula mucilaginosa was reported to contain a 67%
higher carotenoid production than the wild type (Wang, Liu, Yang, &
Wang, 2017). A Chlorella zofingiensis CZ-bkt1 (microalgae) mutant
created using N-methyl-N′-nitro-N-nitrosoguanidine mutagenesis
S. Ram, et al. Journal of Functional Foods 67 (2020) 103867

produces high zeaxanthin (7 ± 0.82 mg g−1), β-carotene

Application on larger volumes faces clogging problems due to sugar and

poor efficiency with non polar/ volatile solvents, thermal degradation,
Toxic by products, non eco-friendly, requires large volumes of solvent

Specific to only those carotenoid having permanent electric moment,

(7.18 ± 0.72 mg g−1) and lutein (13.81 ± 1.23 mg g−1). This mutant

Need to optimize for different samples due to variation in electrical

Not suitable for high water containing samples, low yield of polar
has a defected carotenoid ketolase leading to the accumulation of
zeaxanthin in the thylakoid membrane (Huang, Lin, He, Gong, &
Huang, 2018). Yarrowia lipolytica, an oligeneous yeast, has been re-

Costly, synthesis requires non-eco-friendly chemicals,

ported to contain an adequate supply of precursor compounds and

Non recommended for use in food and feed sectors

High power consumption, added operational costs

energy sources (acetyl-CoA, ATP and NADPH) and is emerging to be a
competitive source for β-carotene production (Matthäus, Ketelhot,
Gatter, & Barth, 2014). Engineered biosynthetic pathways using strong

High equipment and operational costs,

promoters and multiple gene copies have yielded a 100-fold increase in

cis–trans isomerization of carotenoid

β-carotene content than the wild Y. lipolytica strains with a productivity

Optimum conditions requirement

of 4 g L−1 (Gao et al., 2017; Larroude et al., 2018). System metabolic

pectin content of plant matrices

engineering will continue to play central role in the development of
high-performance carotenoid producing strains. Genetic engineering

Expensive instrument,

Prone to degradation
will play a pivotal role in countering high capital investments, thus

DISADVANTAGES
warranting a successful commercialisation.

conductivity

carotenoids

Expensive,
10. Extraction processes for carotenoid production

Basically, carotenoids are lipid-soluble; they are usually extracted

with organic solvents (chloroform, hexane, acetone and petroleum

Low toxicity, minimal use of organic co-solvents

Higher carotenoid yield, higher diffusivity and

Non flammable under wide temperature range

Short extraction time (few minutes), minimal
Suitable for Gram negative bacteria Easy and
ether). The application of non-polar solvents (hexane, petroleum ether

Non thermal, fast extraction, less solvent use

Non-thermal, High extraction efficiency, less

organic solvent utilized, useful for lab-scale

or tetrahydrofuran) for the extraction of non-polar carotene is em-
ployed; and polar solvents (acetone, methanol and ethanol) are a more

No need of sophisticated instruments

suitable choice for polar carotenoid extraction (Saini & Keum, 2018).
The solvent mediated extraction is especially suitable for Gram-nega-
tive bacteria, as they have a densely packed lipid bilayer architecture.

Rapid and highly specific

Environmentally friendly
In recent times, enzyme-mediated extraction processes using lysozyme

Simple and economical

Reduced reaction time

Environmentally safe
energy consumption
have been commonly employed to disintegrate bacterial cell mem-

Renewable source
branes by hydrolyzing the β-1–4-glucosidic bonds in peptidoglycan
ADVANTAGES

permeability,
(Pasquet et al., 2011). Enzymatic extraction improves extraction pro-

experiments
Non toxic
low costs

ficiency, diminishes the use of organic solvents, is environmentally

friendly, renewable and confers specificity. However, enzymes are ex-
pensive, need optimal operating conditions and are prone to degrada-
tion, thus requiring special care during experiments. To overcome low
Uses high voltage pulses induce electroporation of
yield and high extraction costs from enzyme-mediated carotenoid ex-

Disintegrating bacterial cell membrane utilizing

Extraction using supercritical CO2 and ethanol,
Operate at low temperature and high pressure

Operate at low temperature and high pressure

Biomass in contact with organic solvents, cell

traction, a direct and quick extraction is performed using a microwave-

Continuous extraction using organic solvents

cellulotytic enzymes (β-1–4-glucosidic bonds

Cell disruption due to microwave radiation

assisted extraction (MAE) technique. Carotenoid extraction results from

the destruction of cell structures caused due to exposure to microwave
Advantages and disadvantages associated with various carotenoid extraction processes.

radiation. MAE is of two types; microwave-assisted solvent extraction

(MASE) and microwave-assisted solvent-free extraction (MASFE). An-

disintegration by lysozyme)
other sub-type of MASFE, microwave hydro diffusion and gravity
(MHG) has recently gained momentum as it is a green extraction
Use ultrasound waves

method (Nadar, Rao, & Rathod, 2018). However, the MAE application
MODE OF ACTION

is limited to carotenoid molecules having a permanent electric moment

cell membrane

(Butnariu, 2016). Consumer awareness of the associated health issues

disruption

upon exposure to chemically extracted food grade products such as

carotenoids continually leads to advancements in extraction technolo-
gies. Supercritical Fluid Extraction (SFE) is a green alternative for
conventional carotenoid extraction methods. SFE uses solvents gen-
Supercritical and sub critical
Pressurized liquid extraction

Enzyme assisted extraction

Pulsed electric field (PEF)

erally recognised as safe (GRAS) viz., water and carbon dioxide, making
it the most preferred method of extraction in pharmaceuticals and food
fluid extraction (SFE)
Microwave-assisted

Ultrasound-assisted

industries. Additionally, due to its higher permeability and diffusivity

extraction (MAE)

extraction (UAE)

Ionic Liquid (IL)

compared with conventional extraction methods, it enables faster and

Maceration
METHODS

higher carotenoid yields (Nadar et al., 2018). The popular carotenoid

Soxhlet

extraction methods are listed in Table 4 with their pros and cons.
(EAE)
(PLE)

11. Applications of carotenoids

11.1. Carotenoids and human health

Green Measures (GM)
Extraction (ALE)

Extraction (ASE)
Accelerated Solvent
Atmospheric Liquid

Metabolic processes for energy generation occurring in the mi-

tochondrial matrices lead to an ‘oxidative stress environment’ due to
the production of singlet oxygen species and free radicals as by-pro-
Table 4

ducts. These singlet O2 species and free radicals are accountable for a
wide spectrum of conditions such as aging, rheumatoid fever, cancer,

7
S. Ram, et al.
age-related macular degeneration, Alzheimer’s, dementia, ischemic
heart disease and stroke. Carotenoids function as a potent antioxidant
and evade the damaging effects of oxidative stress by quenching free
radicals and singlet O2 species. This antioxidant capability of car-
otenoids is attributed to the presence of their ‘conjugated bonds holding
electron-rich environment’ and the number of double bonds (minimum
9) (Palozza & Krinsky, 1992). An imbalance of antioxidant/reactive
species in the cell causes ‘oxidative stress’, which is the principal con-
tributor to the pathogenesis of chronic diseases.
11.1.1. Carotenoids in cancer prevention
Unlike the classic notion of carotenoids’ primary association with
vitamin A activity, they are now being linked to playing a wider role in
protection and in improving defence mechanisms such as the inhibition
of cell proliferation, the enhancement of cell differentiation, the pro-
tection from photodamage by filtering blue light, the treatment of er-
ythema (skin rashes), the stimulation of cell-to-cell communication, and
in the modulation of carcinogenic metabolism and erythropoietic pro-
toporphyria (EPP), which occurs due to quinidine injection-related
sensitivity by the absorption of shorter light wavelengths, thus resulting
in photosensitivity (Rodriguez-Amaya & Kimura, 2004).
Popular carotenoids such as β-carotene, astaxanthin, lycopene and
zeaxanthin have been reported to demonstrate maximum antioxidant
properties both in vivo and in vitro (Mukherjee, Bose, & Mukhopadhyay,
2017). Several reports have validated the prophylactic effect of a ly-
copene-rich diet as opposed to an oral one (Livny et al., 2002) on
various types of cancers such as bladder (Noviendri, Hasrini, &
Octavianti, 2011), prostate (Rowles, Ranard, Smith, An, & Erdman,
2017), gastric (Franceschi et al., 1994), breast (Dorgan et al., 1998) and
facilitating gap junction communications (Livny et al., 2002). Others
reports also relate carotenoids (α-carotene, lutein, zeaxanthin, lyco-
pene, β-cryptoxanthin, fucoxanthin and astaxanthin) with cancer pre-
vention (Noviendri et al., 2011; Rao & Agarwal, 2000; Vine, Leung, &
Bertram, 2005). β-carotene is the chief carotenoid found in the human
diet, blood and tissue, thus making it the most explored and widely
studied carotenoid out of all of them. Provitamin A activity is attributed
to a beneficial chemotherapeutic effect in the prevention of night
blindness and cancer. However, an inverse relationship has been re-
ported between the occurrence of lung cancer and the plasma levels of
lycopene and β-carotene. For example, heavy smokers and workers
exposed to asbestos were found to be more vulnerable to lung cancers
post β-carotene administration (30 mg day−1) (Goodman et al., 2004).
Another carotenoid, β-cryptoxanthin is reported to invoke a stimulatory
effect on bone formation while exhibiting an antagonistic effect on bone
resorption (the breakdown process of osteoclasts in bone-releasing
minerals). β-cryptoxanthin supplementation may thus be beneficial for
the prevention of osteoporosis, which frequently occurs in post-
menopausal women (Yamaguchi et al., 2006).
Although there is ample evidence of a positive correlation between
carotenoid supplementation and disease prevention, as mentioned
above, some reports also highlight a lack of anticipated benefits, or
even some contradictory results, such as in cases of breast cancer after
carotenoid supplementation (Zhang et al., 1997). This discrepancy is
likely to occur due to a lack of exposure time for supplements needed to
show its preventive benefits in the human body or other experimental
factors, which may lead to an insufficient detection of their beneficial
effects. A lack of definitive evidence linking carotenoids with disease
treatment may also arise due to the complex dynamics that exist be-
tween diet, genetic make-up, the environment and carotenoid vulner-
ability for structural modifications within biological matrices.
11.1.2. Carotenoids and ocular health
Lutein, zeaxanthin and meso-zeaxanthin are found in the human
macula. These carotenoids have the ability to absorb harmful blue light,
thereby protecting the retina from photo damage. These compounds
have attracted much interest in ophthalmic research, especially in the
8
Journal of Functional Foods 67 (2020) 103867
occurrence of age-related macular degeneration (AMD) (Buitendijk,
2018). High lutein/zeaxanthin serum levels show suppressed AMD
conditions. Zeaxanthin is more significantly associated with AMD than
lutein (Sabour-Pickett, Nolan, Loughman, & Beatty, 2012). The Joint
Food and Agriculture Organization/World Health Organization (FAO/
WHO) Expert Committee on Food Additives (JECFA) has established an
Acceptable Daily Intake (ADI) for zeaxanthin for a 0–2 mg kg−1 body
weight (Joint & Additives, 2006).
11.1.3. Carotenoid-mediated multidrug resistance (MDR) reversal
Carotenoids evade the risk of chronic heart disease using various
mechanisms including anti-oxidation properties, the stimulation of
apoptosis of tumor cells and inhibition of oncogene expression
(Garattini, Gianni, & Terao, 2007). The majority of tumor cells express
a multi-drug resistance (MDR) protein belonging to the ATP binding
cassette (ABC) super family. Their expression confers the cancerous
cells with its ability to resist drugs, thereby nullifying the therapeutic
effects of chemotherapy and leading to therapeutic failures. Some stu-
dies have demonstrated that carotenoids are anti-MDR compounds that
act by modulating ABC transporters (Üstün et al., 2002). Apparently,
the altered ABC transporter loses its drug resistance activity (Szakács,
Özvegy, Bakos, Sarkadi, & Váradi, 2000), leading to the induction of
cytotoxicity as conferred by the administered drug, thus reverting the
MDR. Ample reports have proven the beneficial effect of the co-ad-
ministration of carotenoids with chemotherapeutical drugs in ameli-
orating the therapeutic effects of these drugs. For MDR1 gene-trans-
fected mouse lymphoma cells, the most effective compounds were
capsanthin, lycophyll, capsorubin, luteoxanthin-epimers, violeoxanthin
and phytoxanthins, all of which showed a very pronounced antagonistic
effect on the cancer cell lines (Molnár et al., 2006). Rhodamine accu-
mulation in human breast cancer cells MDR/MRP MDA-MB-231 (HTB-
26) showed prominent activity with antheraxanthin, violeoxanthin,
phytoxanthins, lutein and violaxanthin, while the luteoxanthin-epimers
and β-cryptoxanthin showed lesser activity (Molnár et al., 2006). Eid,
El-Readi, and Wink (2012) reported the beneficial effects of car-
otenoids, particularly fucoxanthin in lowering the expression of MDR in
Caco-2 colon cancer cells (Eid et al., 2012).
11.2. Carotenoids as quorum quenchers
Bacterial cells communicate with each other through cell-to-cell
signaling by employing autoinducers or pheromones (acylated homo-
serine lactones and oligo-peptides) via a widely popularised system
known as quorum sensing (QS). The advancement of disease patho-
genicity could be prevented if measures are taken to block QS signaling,
thereby preventing the biofilm genesis (Paluch, Rewak-Soroczyńska,
Jędrusik, Mazurkiewicz, & Jermakow, 2020). The inhibition of a bac-
terial signaling network halts the trigger of the virulence and patho-
genicity of bacterial derived diseases (Cámara, Williams, & Hardman,
2002). There is only one report suggesting the anti-QS potential of
carotenoids using Pseudomonas aeruginosa, a multi-drug resistant pa-
thogen that regulates its virulent gene expressions using QS. Zeax-
anthin, thus reducing the virulence of P. aeruginosa by down-regulating
QS-related gene expression (Gökalsın, Aksoydan, Erman, & Sesal,
2017).
11.3. Carotenoids and animal nutrition
11.3.1. Carotenoids as feed additives
The increase in the requirement of animal products as meat and
eggs for human consumption has been accompanied by an upsurge in
the production of improved poultry in terms of higher quality products
(Moreno, Díaz-Gómez, Nogareda, Angulo, Sandmann, Portero-Otin,
Serrano, Twyman, Capell, Zhu, & Christou, 2016). A cocktail of car-
otenoids (astaxanthin, β-carotene, lutein) as an aquafeed are provided
for animals, fish and birds to deepen their flesh and egg colouration
S. Ram, et al.
Fig. 3. SWOT analysis of bacterial carotenoids.
through accumulation (Breitenbach et al., 2016; Nogareda et al., 2016;
Nogueira et al., 2017). Supplementing aquafeed with carotenoids is
pertinent to augmenting the quality of animal products, thereby making
the products nutritionally enhanced and economically beneficial, thus
satisfying for the consumers and profitable for the producers.
11.3.2. Carotenoids as a flavour enhancer, food pigment and allelochemical
Food colour determines consumer acceptability and desirability.
Humans perceive the appearance of their food as an index for food
quality. Carotenoids such as β- carotene, β-apo-8′-carotenal and can-
thaxanthin are used to impart colour to many processed foods (e.g.
butter, dairy products, margarine, confectionery and baked foods).
Besides playing an important role as functional food ingredients, car-
otenoids also are precursors for some popular volatile flavouring
compounds known as apocarotenoids (Cataldo, López, Cárcamo, &
Agosin, 2016). Reports suggest that carotenoid supplementation is not
associated with its nutritive value, but its beneficial role is in the pre-
vention or delaying the onset of chronic diseases. The oxidation of
carotenoids yields C13-C11-C10 and C9 derivatives, which constitute the
aroma compounds responsible for providing flavour and odour. For
example, β-ionone, β-damascenone and saframal are widely found in
roses, black tea and raspberries. Crocetin, derived from saffron (the
crocus flower) is an apocarotenoid without provitamin A activity
(Giaccio, 2004). Crocetin has been demonstrated to have a significant
beneficial effect on coronary heart disease by inhibiting lipoprotein
oxidation (Devivasha & Javadi, 2018; Giaccio, 2004), on neurodegen-
erative diseases (Finley & Gao, 2017), on retinal function recovery by
increasing retinal blood flow (Sepahi et al., 2018), on reducing blood
9
Journal of Functional Foods 67 (2020) 103867
pressure and maintaining kidney function (Khorasany & Hosseinzadeh,
2016), as well as being used as an anticonvulsive agent (Hosseinzadeh
& Younesi, 2002). The most notable discovery in relation to crocetin is
the enhanced oxygen diffusivity during shock and reperfusion, which
ultimately leads to increased ATP production (Dhar et al., 2005).
12. SWOT (Strengths, Weaknesses, Opportunities, and Threats)
analysis
A SWOT analysis is conducted to perceive the significance of the
four major components (Strengths, Weaknesses, Opportunities and
Threats) to achieve the economic viability of carotenoid productions
from bacterial sources (Fig. 3).
12.1. Strengths
The bacterial carotenoid is pursued due to its natural origin, its
varied shades of colour, its low media requirements and its lack of
seasonal dependency. The co-production of other valuable compounds
such as enzymes, amino acids, vitamins etc. also correspond to value
addition. The use of agro-industrial waste such as sugarcane and beet
molasses such as low-cost carbon and nitrogen sources further reduces
carotenoid manufacturing costs and enabling the safe disposal of waste.
12.2. Weakness
The high costs associated with the production of bacterial car-
otenoids hinders their commercial value and lowers their market share.
S. Ram, et al.
Low carotenoid content in bacteria can be resolved by using process
optimisation strategies for making it more economical.
12.3. Opportunities
Consumer attraction towards natural carotenoids and environ-
mental safety has swapped interest in natural to synthetic sources.
Additionally, the prokaryotic nature of bacteria enables easier genetic
modifications yielding hyper-producing strains. Since only < 1% of
the bacterial population is known, exploration of other bacterial species
may lead to the identification of novel and rare carotenoids.
12.4. Threats:
Competition from already existing natural commercial sources of
carotenoid-like microalgae and plant, present challenges for the ac-
ceptability of bacterial-derived carotenoids. The non-competition of
bacterial cultivation methods with arable land makes them a superior
alternate, since upstream production will not compete with crop plants.
Also, strain improvement through genetic engineering can tackle the
low yields in promising strains.
13. Present challenges and future directions
A plethora of territories are still understudied and unexplored,
which makes a profusely diverse spectrum of compounds yet to be
identified. A thorough exploration expedition needs to be undertaken
with multifarious specialists from interdisciplinary origins. Stimulating
carotenogenesis using a two-stage starvation strategy has long been
employed in microalgae and recently in some yeasts for accumulating
carotenoids. This could be an effective approach to obtaining high-yield
carotenoid-producing bacteria and to revealing entirely novel car-
otenoid structures. A complete knowledge involving genetic and me-
tabolon studies of a complex intertwined biosynthetic pathway is
needed to establish preventive actions for many chronic diseases. This
insight will be helpful in creating novel drugs for the prevention of
many lethal diseases. Developing defective mutants could pave the way
for the construction of industrially relevant strains. Nevertheless, any
genetically modified strain must be scrutinised for its use as a dietary
inclusion, for its cost-effective carotenoid production and for its lack of
any harmful impact on the environment.
Interdisciplinary advancements involving microbiologists, mole-
cular biologists, chemical analysts, fermentation technologists, extrac-
tion specialists and toxicologists must be trained further in order to
construct a high impact and commercially relevant carotenoid source.
Declaration of Competing Interest
We declare that we have no conflict of interest.
Acknowledgments
This manuscript has been assigned by CSIR-CSMCRI registration
number of CSIR-CSMCRI – 139/2018. SR, FS and MM would like to
acknowledge CSIR for the SRF fellowship and SRT would like to thank
UGC-NFHE for their financial support. SR, MM and SRT are grateful to
AcSIR for their PhD enrolment. FS is thankful to Bhavnagar University
for PhD enrolment.
Appendix A. Supplementary material
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.jff.2020.103867.
10
Journal of Functional Foods 67 (2020) 103867
References
Al-Babili, S., & Bouwmeester, H. J. (2015). Strigolactones, a novel carotenoid-derived
plant hormone. Annual Review of Plant Biology, 66, 161–186. https://doi.org/10.
1146/annurevarplant-043014-114759.
Alper, H., Miyaoku, K., & Stephanopoulos, G. (2006). Characterization of lycopene-
overproducing E. coli strains in high cell density fermentations. Applied Microbiology
and Biotechnology, 72(5), 968–974. https://doi.org/10.1007/s00253-006-0357-y.
Antognoni, F., Mandrioli, R., Potente, G., Saa, D. L. T., & Gianotti, A. (2019). Changes in
carotenoids, phenolic acids and antioxidant capacity in bread wheat doughs fer-
mented with different lactic acid bacteria strains. Food Chemistry, 292, 211–216.
https://doi.org/10.1016/j.foodchem.2019.04.061.
Aruldass, C. A., Dufossé, L., & Ahmad, W. A. (2018). Current perspective of yellowish-
orange pigments from microorganisms-a review. Journal of Cleaner Production, 180,
168–182. https://doi.org/10.1016/j.jclepro.2018.01.093.
Aryee, A. N., Agyei, D., & Akanbi, T. O. (2018). Recovery and utilization of seaweed
pigments in food processing. Current Opinion in Food Science. https://doi.org/10.
1016/j.cofs.2018.03.013.
Asker, D., Awad, T. S., Beppu, T., & Ueda, K. (2018). Screening and profiling of natural
ketocarotenoids from environmental aquatic bacterial isolates. Food Chemistry, 253,
247–254. https://doi.org/10.1016/j.foodchem.2018.01.066.
Asker, D., Awad, T. S., Beppu, T., & Ueda, K. (2012). Isolation, characterization, and di-
versity of novel radiotolerant carotenoid-producing bacteria Microbial Carotenoids from
Bacteria and Microalgae. Totowa, NJ: Springer. Humana Press 10.1007/978-1-61779-
879-5_3 21 60.
Asker, D., Beppu, T., & Ueda, K. (2007). Mesoflavibacter zeaxanthinifaciens gen. nov., sp.
nov., a novel zeaxanthin-producing marine bacterium of the family
Flavobacteriaceae. Systematic and Applied Microbiology, 30(4), 291–296. https://doi.
org/10.1016/j.syapm.2006.12.003.
Asker, D., Beppu, T., & Ueda, K. (2007). Zeaxanthinibacter enoshimensis gen. nov., sp. nov.,
a novel zeaxanthin-producing marine bacterium of the family Flavobacteriaceae,
isolated from seawater off Enoshima Island, Japan. International Journal of Systematic
and Evolutionary Microbiology, 57(4), 837–843. https://doi.org/10.1099/ijs.0.
64682-0.
Batghare, A. H., Singh, N., & Moholkar, V. S. (2018). Investigations in ultra-
sound–induced enhancement of astaxanthin production by wild strain Phaffia rho-
dozyma MTCC 7536. Bioresource Technology, 254, 166–173. https://doi.org/10.1016/
j.biortech.2018.01.073.
Ben-Amotz, A., & Levy, Y. (1996). Bioavailability of a natural isomer mixture compared
with synthetic all-trans beta-carotene in human serum. The American Jjournal of
Clinical Nutrition, 63(5), 729–734. https://doi.org/10.1093/ajcn/63.5.729.
Beuttler, H., Hoffmann, J., Jeske, M., Hauer, B., Schmid, R. D., Altenbuchner, J., &
Urlacher, V. B. (2011). Biosynthesis of zeaxanthin in recombinant Pseudomonas pu-
tida. Applied Microbiology and Biotechnology, 89(4), 1137–1147. https://doi.org/10.
1007/s00253-010-2961-0.
Bhosale, P. (2004). Environmental and cultural stimulants in the production of car-
otenoids from microorganisms. Applied Microbiology and Biotechnology, 63(4)
351–361.https: //doi.org/10.1007/s00253-003-1441-1.
Breitenbach, J., Nogueira, M., Farré, G., Zhu, C., Capell, T., Christou, P., ... Sandmann, G.
(2016). Engineered maize as a source of astaxanthin: Processing and application as
fish feed. Transgenic Research, 25(6), 785–793. https://doi.org/10.1007/s11248-016-
9971-3.
Britton, G., Liaaen-Jensen, S., & Pfander, H. (Eds.). (1995). Carotenoids: Spectroscopy.
Vol. 1B. Birkhäuser Verlag.
Buitendijk, G. (2018). Age-related Macular Degeneration: from risk profiles towards
prediction models.
Butnariu, M. (2016). Methods of analysis (extraction, separation, identification and
quantification) of carotenoids from natural products. Journal of Ecosystem and
Ecography, 6(2), https://doi.org/10.4172/2157-7625.1000193.
Cámara, M., Williams, P., & Hardman, A. (2002). Controlling infection by tuning in and
turning down the volume of bacterial small-talk. The Lancet Infectious Diseases, 2(11),
667–676. https://doi.org/10.1016/S1473-3099(02)00447-4.
Cataldo, V. F., López, J., Cárcamo, M., & Agosin, E. (2016). Chemical vs. biotechnological
synthesis of C 13-apocarotenoids: Current methods, applications and perspectives.
Applied Microbiology and Biotechnology, 100(13), 5703–5718. https://doi.org/10.
1007/s00253-016-7583-8.
Chattopadhyay, M., Jagannadham, M., Vairamani, M., & Shivaji, S. (1997). Carotenoid
pigments of an antarctic psychrotrophic bacterium Micrococcus roseus: Temperature
dependent biosynthesis, structure, and interaction with synthetic membranes.
Biochemical and Biophysical Research Communications, 239(1), 85–90. https://doi.org/
10.1006/bbrc.1997.7433.
Chen, F., Li, H.-B., Wong, R. N.-S., Ji, B., & Jiang, Y. (2005). Isolation and purification of
the bioactive carotenoid zeaxanthin from the microalga Microcystis aeruginosa by
high-speed counter-current chromatography. Journal of Chromatography A, 1064(2),
183–186. https://doi.org/10.1016/j.chroma.2004.12.065.
Chen, Y., Xie, B., Yang, J., Chen, J., & Sun, Z. (2018). Identification of microbial car-
otenoids and isoprenoid quinones from Rhodococcus sp. B7740 and its stability in the
presence of iron in model gastric conditions. Food Chemistry, 240, 204–211. https://
doi.org/10.1016/j.foodchem.2017.06.067.
Choi, H. S., Lee, S. Y., Kim, T. Y., & Woo, H. M. (2010). In silico identification of gene
amplification targets for improvement of lycopene production. Applied and
Environmental Microbiology. 76(10), 3097–3105. https://doi.org/10.1128/AEM.
00115-10.
Christian, D., Zhang, J., Sawdon, A. J., & Peng, C.-A. (2018). Enhanced astaxanthin ac-
cumulation in Haematococcus pluvialis using high carbon dioxide concentration and
S. Ram, et al.
light illumination. Bioresource Technology, 256, 548–551. https://doi.org/10.1016/j.
biortech.2018.02.074.
D’Souza, S. E., Altekar, W., & D’Souza, S. (1997). Adaptive response of Haloferax medi-
terranei to low concentrations of NaCl (< 20%) in the growth medium. Archives of
Microbiology, 168(1), 68–71. https://doi.org/10.1007/s002030050471.
Deinove (n.d.). Carotenoids market. Natural carotenoids meeting consumer needs.
< http://www.deinove.com/en/profile/strategy-and-markets/carotenoids-market/
> Accessed 20 May 2019.
Devivasha, B., & Javadi, M. (2018). Cancer Cell chemoresistance and chemosensitization.
World Scientific.
Dhar, A., Cherian, G., Dhar, G., Ray, G., Sharma, R., & Banerjee, S. K. (2005). Molecular
basis of protective effect by crocetin on survival and liver tissue damage following
hemorrhagic shock. Molecular and Cellular Biochemistry, 278(1–2), 139–146. https://
doi.org/10.1007/s11010-005-7155-1.
Dorgan, J. F., Sowell, A., Swanson, C. A., Potischman, N., Miller, R., Schussler, N., &
Stephenson, H. E. (1998). Relationships of serum carotenoids, retinol, α-tocopherol,
and selenium with breast cancer risk: Results from a prospective study in Columbia,
Missouri (United States). Cancer Causes & Control, 9(1), 89–97. https://doi.org/10.
1023/A:1008857521992.
Dufossé, L., Galaup, P., Yaron, A., Arad, S. M., Blanc, P., Murthy, K. N. C., & Ravishankar,
G. A. (2005). Microorganisms and microalgae as sources of pigments for food use: A
scientific oddity or an industrial reality? Trends in Food Science & Technology, 16(9),
389–406. https://doi.org/10.1016/j.tifs.2005.02.006.
Eid, S. Y., El-Readi, M. Z., & Wink, M. (2012). Carotenoids reverse multidrug resistance in
cancer cells by interfering with ABC-transporters. Phytomedicine, 19(11), 977–987.
https://doi.org/10.1016/j.phymed.2012.05.010.
Fang, C.-J., Ku, K.-L., Lee, M.-H., & Su, N.-W. (2010). Influence of nutritive factors on C50
carotenoids production by Haloferax mediterranei ATCC 33500 with two-stage culti-
vation. Bioresource Technology, 101(16), 6487–6493. https://doi.org/10.1016/j.
biortech.2010.03.044.
Farmer, W. R., & Liao, J. C. (2001). Precursor balancing for metabolic engineering of
lycopene production in Escherichia coli. Biotechnology Progress, 17(1), 57–61. https://
doi.org/10.1021/bp000137t.
Fasano, E., Serini, S., Mondella, N., Trombino, S., Celleno, L., Lanza, P., Cittadini, A., &
Calviello, G. (2014). Antioxidant and Anti-Inflammatory Effects of Selected Natural
Compounds Contained in a Dietary Supplement on Two Human Immortalized
Keratinocyte Lines. BioMed Research International, 2014, 1–11. https://doi.org/10.
1155/2014/327452.
Finley, J. W., & Gao, S. (2017). A perspective on Crocus sativus L. (Saffron) constituent
crocin: A potent water-soluble antioxidant and potential therapy for Alzheimer’s
disease. Journal of Agricultural and Food Chemistry, 65(5), 1005–1020. https://doi.
org/10.1021/acs.jafc.6b04398.
Flores-Cotera, L., Martin, R., & Sanchez, S. (2001). Citrate, a possible precursor of as-
taxanthin in Phaffia rhodozyma: Influence of varying levels of ammonium, phosphate
and citrate in a chemically defined medium. Applied Microbiology and Biotechnology,
55(3), 341–347. https://doi.org/10.1007/s002530000498.
Fong, N., Burgess, M., Barrow, K., & Glenn, D. (2001). Carotenoid accumulation in the
psychrotrophic bacterium Arthrobacter agilis in response to thermal and salt stress.
Applied Microbiology and Biotechnology, 56(5–6), 750–756. https://doi.org/10.1007/
s002530100739.
Fontes, M., Ruiz-Vázquez, R., & Murillo, F. J. (1993). Growth phase dependence of the
activation of a bacterial gene for carotenoid synthesis by blue light. The EMBO
Journal, 12(4), 1265–1275. https://doi.org/10.1002/j.1460-2075.1993.tb05771.x.
Franceschi, S., Bidoli, E., Vecchia, C. L., Talamini, R., D'Avanzo, B., & Negri, E. (1994).
Tomatoes and risk of digestive-tract cancers. International Journal of Cancer, 59(2),
181–184. https://doi.org/10.1002/ijc.2910590207.
Ganapathy, A., Jayavel, S., & Natesan, S. (2016). Draft Genome Sequence of Carotenoid
Producing Yellow Pigmented Planococcus maritimus MKU009. J. Genomics, 4, 23–25.
https://doi.org/10.7150/jgen.15533.
Gao, S., Tong, Y., Zhu, L., Ge, M., Zhang, Y., Chen, D., ... Yang, S. (2017). Iterative in-
tegration of multiple-copy pathway genes in Yarrowia lipolytica for heterologous β-
carotene production. Metabolic Engineering, 41, 192–201. https://doi.org/10.1016/j.
ymben.2017.04.004.
Garattini, E., Gianni, M., & Terao, M. (2007). Retinoids as differentiating agents in on-
cology: A network of interactions with intracellular pathways as the basis for rational
therapeutic combinations. Current Pharmaceutical Design, 13(13), 1375–1400.
https://doi.org/10.2174/138161207780618786.
Giaccio, M. (2004). Crocetin from saffron: An active component of an ancient spice.
Critical Reviews in Food Science and Nutrition, 44(3), 155–172. https://doi.org/10.
1080/10408690490441433.
Gharibzahedi, S. M. T., Razavi, S. H., Mousavi, S. M., & Moayedi, V. (2012). High effi-
ciency canthaxanthin production by a novel mutant isolated from Dietzia na-
tronolimnaea HS-1 using central composite design analysis. Industrial Crops and
Products, 40, 345–354. https://doi.org/10.1016/j.indcrop.2012.03.030.
Gökalsın, B., Aksoydan, B., Erman, B., & Sesal, N. C. (2017). Reducing virulence and
biofilm of Pseudomonas aeruginosa by potential quorum sensing inhibitor carotenoid:
Zeaxanthin. Microbial Ecology, 74(2), 466–473. https://doi.org/10.1007/s00248-
017-0949-3.
Goodman, G. E., Thornquist, M. D., Balmes, J., Cullen, M. R., Meyskens, F. L., Jr, Omenn,
G. S., ... Williams, J. H., Jr (2004). The Beta-Carotene and Retinol Efficacy Trial:
Incidence of lung cancer and cardiovascular disease mortality during 6-year follow-
up after stopping β-carotene and retinol supplements. Journal of the National Cancer
Institute, 96(23), 1743–1750. https://doi.org/10.1093/jnci/djh320.
Group, E. D. C. C. S. (1993). Antioxidant status and neovascular age-related macular
degeneration. Archives of Opthalmology, 111, 104–109.
Guyomarc’h, F., Binet, A., & Dufossé, L. (2000). Production of carotenoids by
11
Journal of Functional Foods 67 (2020) 103867
Brevibacterium linens: Variation among strains, kinetic aspects and HPLC profiles.
Journal of Industrial Microbiology and Biotechnology, 24(1), 64–70. https://doi.org/10.
1038/sj.jim.2900761.
Hameed, A., Shahina, M., Lin, S.-Y., Lai, W.-A., Hsu, Y.-H., Liu, Y.-C., & Young, C.-C.
(2014). Aquibacter zeaxanthinifaciens gen. nov., sp. nov., a zeaxanthin-producing
bacterium of the family Flavobacteriaceae isolated from surface seawater, and
emended descriptions of the genera Aestuariibaculum and Gaetbulibacter.
International Journal of Systematic and Evolutionary Microbiology, 64(1), 138–145.
https://doi.org/10.1099/ijs.0.052621-0.
Henke, N. A., Wiebe, D., Pérez-García, F., Peters-Wendisch, P., & Wendisch, V. F. (2018).
Coproduction of cell-bound and secreted value-added compounds: Simultaneous
production of carotenoids and amino acids by Corynebacterium glutamicum.
Bioresource Technology, 247, 744–752. https://doi.org/10.1016/j.biortech.2017.09.
167.
Hosseinzadeh, H., & Younesi, H. M. (2002). Antinociceptive and anti-inflammatory effects
of Crocus sativus L. stigma and petal extracts in mice. BMC Pharmacology, 2(1), 7.
https://doi.org/10.1186/1471-2210-2-7.
Huang, W., Lin, Y., He, M., Gong, Y., & Huang, J. (2018). Induced high-yield production
of zeaxanthin, lutein, and β-carotene by a mutant of Chlorella zofingiensis. Journal of
Agricultural and Food Chemistry, 66(4), 891–897. https://doi.org/10.1021/acs.jafc.
7b05400.
Iigusa, H., Yoshida, Y., & Hasunuma, K. (2005). Oxygen and hydrogen peroxide enhance
light-induced carotenoid synthesis in Neurospora crassa. FEBS Letters, 579(18),
4012–4016. https://doi.org/10.1016/j.febslet.2005.06.014.
Joint, F., & Additives, W. E. C. o. F. (2006). Safety evaluation of certain food additives.
Jung, J., Lim, J. H., Kim, S. Y., Im, D.-K., Seok, J. Y., Lee, S.-J. V., ... Jung, G. Y. (2016).
Precise precursor rebalancing for isoprenoids production by fine control of gapA
expression in Escherichia coli. Metabolic Engineering, 38, 401–408. https://doi.org/10.
1016/j.ymben.2016.10.003.
Kim, W. J., Kim, H. U., & Lee, S. Y. (2017). Current state and applications of microbial
genome-scale metabolic models. Current Opinion in Systems Biology, 2, 10–18. https://
doi.org/10.1016/j.coisb.2017.03.001.
Kim, S. H., Kim, J. H., Lee, B. Y., & Lee, P. C. (2014). The astaxanthin dideoxyglycoside
biosynthesis pathway in Sphingomonas sp. PB304. Applied Microbiology and
Biotechnology, 98(24), 9993–10003. https://doi.org/10.1007/s00253-014-6050-7.
Kirti, K., Amita, S., Priti, S., Mukesh Kumar, A., & Jyoti, S. (2014). Colorful world of
microbes: Carotenoids and their applications. Advances in Biology, 2014. https://doi.
org/10.1155/2014/837891.
Khorasany, A. R., & Hosseinzadeh, H. (2016). Therapeutic effects of saffron (Crocus sativus
L.) in digestive disorders: A review. Iranian Journal of Basic Medical Sciences, 19(5),
455.
Kot, A. M., Błażejak, S., Gientka, I., Kieliszek, M., & Bryś, J. (2018). Torulene and tor-
ularhodin:“new” fungal carotenoids for industry? Microbial Cell Factories, 17(1), 49.
https://doi.org/10.1186/s12934-018-0893-z.
Lagarde, D., Beuf, L., & Vermaas, W. (2000). Increased production of zeaxanthin and
other pigments by application of genetic engineering techniques to Synechocystis sp.
strain PCC 6803. Applied and Environmental Microbiolog, 66(1), 64–72. https://doi.
org/10.1128/AEM.66.1.64-72.2000.
Larroude, M., Celinska, E., Back, A., Thomas, S., Nicaud, J. M., & Ledesma-Amaro, R.
(2018). A synthetic biology approach to transform Yarrowia lipolytica into a compe-
titive biotechnological producer of β-carotene. Biotechnology and Bioengineering,
115(2), 464–472. https://doi.org/10.1002/bit.26473.
Lee, P. C., Mijts, B. N., & Schmidt-Dannert, C. (2004). Investigation of factors influencing
production of the monocyclic carotenoid torulene in metabolically engineered
Escherichia coli. Applied Microbiology and Biotechnology, 65(5), 538–546. https://doi.
org/10.1007/s00253-004-1619-1.
Li, X.-R., Tian, G.-Q., Shen, H.-J., & Liu, J.-Z. (2015). Metabolic engineering of Escherichia
coli to produce zeaxanthin. Journal of Industrial Microbiology & Biotechnology, 42(4),
627–636. https://doi.org/10.1007/s10295-014-1565-6.
Livny, O., Kaplan, I., Reifen, R., Polak-Charcon, S., Madar, Z., & Schwartz, B. (2002).
Lycopene inhibits proliferation and enhances gap-junction communication of KB-1
human oral tumor cells. The Journal of Nutrition, 132(12), 3754–3759. https://doi.
org/10.1093/jn/132.12.3754.
Lu, Q., Bu, Y.-F., & Liu, J. -Z. (2017). Metabolic engineering of Escherichia coli for pro-
ducing astaxanthin as the predominant carotenoid. Marine Drugs, 15(10), 296.
https://doi.org/ 10.3390/md15100296.
Lorquin, J., Molouba, F., & Dreyfus, B. L. (1997). Identification of the carotenoid pigment
canthaxanthin from photosynthetic Bradyrhizobium strains. Applied and Environmental
Microbiology, 63(3), 1151–1154. https://doi.org/10.1128/AEM.63.3.1151-1154.
1997.
Lutnaes, B. F., Strand, Å., Pétursdóttir, S. K., & Liaaen-Jensen, S. (2004). Carotenoids of
thermophilic bacteria—Rhodothermus marinus from submarine Icelandic hot springs.
Biochemical Systematics and Ecology, 32(5), 455–468. https://doi.org/10.1016/j.bse.
2003.09.005.
Lv, X., Gu, J., Wang, F., Xie, W., Liu, M., Ye, L., & Yu, H. (2016). Combinatorial pathway
optimization in Escherichia coli by directed co-evolution of rate-limiting enzymes and
modular pathway engineering. Biotechnology and Bioengineering, 113(12), 2661–2669.
https://doi.org/10.1002/bit.26034.
Market Research Report. Carotenoids Market by Type (Astaxanthin, Beta-Carotene,
Canthaxanthin, Lutein, Lycopene, & Zeaxanthin), Source (Synthetic and Natural),
Application (Supplements, Food, Feed, and Cosmetics), & by Region - Global Trends
& Forecasts to 2021. (2016) < https://www.marketsandmarkets.com/Market-
Reports/carotenoid-market158421566.html/ > Accessed 20 May 2019.
Matthäus, F., Ketelhot, M., Gatter, M., & Barth, G. (2014). Production of lycopene in the
non-carotenoid-producing yeast Yarrowia lipolytica. Applied and Environmental
Microbiology, 80(5), 1660–1669. https://doi.org/10.1128/AEM.03167-13.
S. Ram, et al.
McWilliams, A. (2018). The global market for carotenoids. BCC Research: Wellesley, MA,
USA.
Molnár, J., Gyémánt, N., Tanaka, M., Hohmann, J., Bergmann-Leitner, E., Molnár, P., ...
Ferreira, M. J. (2006). Inhibition of multidrug resistance of cancer cells by natural
diterpenes, triterpenes and carotenoids. Current Pharmaceutical Design, 12(3),
287–311. https://doi.org/10.2174/138161206775201893.
Montero-Lobato, Z., Ramos-Merchante, A., Fuentes, J. L., Sayago, A., Fernández-
Recamales, Á., Martínez-Espinosa, R. M., ... Garbayo, I. (2018). Optimization of
growth and carotenoid production by Haloferax mediterranei using response surface
methodology. Marine Drugs, 16(10), 372. https://doi.org/10.3390/md16100372.
Moran, N. A., & Jarvik, T. (2010). Lateral transfer of genes from fungi underlies car-
otenoid production in aphids. Science, 328(5978), 624–627. https://doi.org/10.
1126/science.1187113.
Moreno, J. A., Díaz-Gómez, J., Nogareda, C., Angulo, E., Sandmann, G., Portero-Otin, M.,
Serrano, J. C., Twyman, R. M., Capell, T., Zhu, C., & Christou, P. (2016). The dis-
tribution of carotenoids in hens fed on biofortified maize is influenced by feed
composition, absorption, resource allocation and storage. Scientific Reports, 6,
35346. https://doi.org/ 10.1038/srep35346 (2016).
Mukherjee, T., Bose, S., & Mukhopadhyay, S. K. (2017). Antioxidant properties of the
carotenoid extracts of three Deinococcus-Thermus phylum bacteria, Meiothermus sp.
strains RP and TP and Thermus sp. strain YY from Paniphala hot spring, India. Nutrire,
42(1), 7. https://doi.org/10.1186/s41110-017-0032-3.
Nadar, S. S., Rao, P., & Rathod, V. K. (2018). Enzyme assisted extraction of biomolecules
as an approach to novel extraction technology: A review. Food Research International,
108, 309–330. https://doi.org/10.1016/j.foodres.2018.03.006.
Neagu, S., Cojoc, R., Enache, M., Mocioiu, O. C., Precupas, A., Popa, V. T., ... Enache, M.
(2019). Biotransformation of Waste Glycerol from Biodiesel Industry in Carotenoids
Compounds by Halophilic Microorganisms. Waste and Biomass Valorization, 10(1),
45–52. https://doi.org/10.1007/s12649-017-0040-9.
Nogareda, C., Moreno, J. A., Angulo, E., Sandmann, G., Portero, M., Capell, T., ...
Christou, P. (2016). Carotenoid-enriched transgenic corn delivers bioavailable car-
otenoids to poultry and protects them against coccidiosis. Plant Biotechnology Journal,
14(1), 160–168. https://doi.org/10.1111/pbi.12369.
Nogueira, M., Enfissi, E. M., Valenzuela, M. E. M., Menard, G. N., Driller, R. L., Eastmond,
P. J., ... Fraser, P. D. (2017). Engineering of tomato for the sustainable production of
ketocarotenoids and its evaluation in aquaculture feed. Proceedings of the National
Academy of Sciences, 114(41), 10876–10881. https://doi.org/10.1073/pnas.
1708349114.
Noviendri, D., Hasrini, R. F., & Octavianti, F. (2011). Carotenoids: Sources, medicinal
properties and their application in food and nutraceutical industry. Journal of
Medicinal Plants Research, 5(33), 7119–7131. https://doi.org/10.5897/JMPRx11.
011.
Numan, M., Bashir, S., Mumtaz, R., Tayyab, S., Rehman, N. U., Khan, A. L., ... Al-Harrasi,
A. (2018). Therapeutic applications of bacterial pigments: A review of current status
and future opportunities. 3. Biotech, 8(4), 207. https://doi.org/10.1007/s13205-018-
1227-x.
Oren, A. (2009). Microbial diversity and microbial abundance in salt-saturated brines:
Why are the waters of hypersaline lakes red? Natural Resources and Environmental
Issues, 15(1), 49.
Osanjo, G. O., Muthike, E. W., Tsuma, L., Okoth, M. W., Bulimo, W. D., Luuml, H., ...
Golyshin, P. N. (2009). A salt lake extremophile, Paracoccus bogoriensis sp. nov., ef-
ficiently produces xanthophyll carotenoids. African Journal of Microbiology Research,
3(8), 426–433. https://doi.org/10.5897/AJMR.
Paliwal, C., Mitra, M., Bhayani, K., Bharadwaj, S. V., Ghosh, T., Dubey, S., & Mishra, S.
(2017). Abiotic stresses as tools for metabolites in microalgae. Bioresource Technology.
https://doi.org/10.1016/j.biortech.2017.05.058.
Paliwal, C., Pancha, I., Ghosh, T., Maurya, R., Chokshi, K., Bharadwaj, S. V., ... Mishra, S.
(2015). Selective carotenoid accumulation by varying nutrient media and salinity in
Synechocystis sp. CCNM 2501. Bioresource Technology, 197, 363–368. https://doi.org/
10.1016/j.biortech.2015.08.122.
Palozza, P., & Krinsky, N. I. (1992). [38] Antioxidant effects of carotenoids in Vivo and in
Vitro: An overview. Methods in enzymology: Vol. 213, (pp. 403–420). Academic Press.
https://doi.org/10.1016/0076-6879(92)13142-K.
Paluch, E., Rewak-Soroczyńska, J., Jędrusik, I., Mazurkiewicz, E., & Jermakow, K. (2020).
Prevention of biofilm formation by quorum quenching. Applied Microbiology and
Biotechnology, 1–11. https://doi.org/10.1007/s00253-020-10349-w.
Park, S. Y., Binkley, R. M., Kim, W. J., Lee, M. H., & Lee, S. Y. (2018). Metabolic en-
gineering of Escherichia coli for high-level astaxanthin production with high pro-
ductivity. Metabolic Engineering, 49, 105–115. https://doi.org/10.1016/j.ymben.
2018.08.002.
Pasquet, V., Chérouvrier, J.-R., Farhat, F., Thiéry, V., Piot, J.-M., Bérard, J. B., ... Cadoret,
J.-P. (2011). Study on the microalgal pigments extraction process: Performance of
microwave assisted extraction. Process Biochemistry, 46(1), 59–67. https://doi.org/
10.1016/j.procbio.2010.07.009.
Patthawaro, S., Lomthaisong, K., & Saejung, C. (2019). Bioconversion of Agro-Industrial
Waste to Value-Added Product Lycopene by Photosynthetic Bacterium
Rhodopseudomonas faecalis and Its Carotenoid Composition. Waste and Biomass
Valorization, 1–12. https://doi.org/10.1007/s12649-018-00571-z.
Pénicaud, C., Achir, N., Dhuique-Mayer, C., Dornier, M., & Bohuon, P. (2011).
Degradation of β-carotene during fruit and vegetable processing or storage: Reaction
mechanisms and kinetic aspects: A review. Fruits, 66(6), 417–440. https://doi.org/
10.1051/fruits/2011058.
Pfleger, B. F., Pitera, D. J., Smolke, C. D., & Keasling, J. D. (2006). Combinatorial en-
gineering of intergenic regions in operons tunes expression of multiple genes. Nature
Biotechnology, 24(8), 1027–1032. https://doi.org/10.1038/nbt1226.
Ram, S., Paliwal, C., & Mishra, S. (2019). Growth medium and nitrogen stress sparked
12
Journal of Functional Foods 67 (2020) 103867
biochemical and carotenogenic alterations in Scenedesmus sp. CCNM 1028.
Bioresource Technology Reports, 100194. https://doi.org/10.1016/j.biteb.2019.
100194.
Rao, A. V., & Agarwal, S. (2000). Role of antioxidant lycopene in cancer and heart dis-
ease. Journal of the American College of Nutrition, 19(5), 563–569. https://doi.org/10.
1080/07315724.2000.10718953.
Rodriguez-Amaya, D. B., & Kimura, M. (2004). HarvestPlus handbook for carotenoid ana-
lysis (Vol. 2). Washington, DC: International Food Policy Research Institute (IFPRI).
Rodrıguez-Concepción, M., & Boronat, A. (2002). Elucidation of the methylerythritol
phosphate pathway for isoprenoid biosynthesis in bacteria and plastids. A metabolic
milestone achieved through genomics. Plant Physiology, 130(3), 1079–1089. https://
doi.org/10.1104/pp.007138.
Rodríguez-Villalón, A., Pérez-Gil, J., & Rodríguez-Concepción, M. (2008). Carotenoid
accumulation in bacteria with enhanced supply of isoprenoid precursors by upre-
gulation of exogenous or endogenous pathways. Journal of Biotechnology, 135(1),
78–84. https://doi.org/10.1016/j.jbiotec.2008.02.023.
Rohmer, M., Knani, M., Simonin, P., Sutter, B., & Sahm, H. (1993). Isoprenoid bio-
synthesis in bacteria: A novel pathway for the early steps leading to isopentenyl di-
phosphate. Biochemical Journal, 295(2), 517–524. https://doi.org/10.1042/
bj2950517.
Rojas-Garbanzo, C., Gleichenhagen, M., Heller, A., Esquivel, P., Schulze-Kaysers, N., &
Schieber, A. (2017). Carotenoid profile, antioxidant capacity, and chromoplasts of
pink guava (Psidium guajava L. Cv. ‘Criolla’) during fruit ripening. Journal of
Agricultural and Food Chemistry, 65(18), 3737–3747. https://doi.org/10.1021/acs.
jafc.6b04560.
Rowles, J., Ranard, K., Smith, J., An, R., & Erdman, J. W. (2017). Increased dietary and
circulating lycopene are associated with reduced prostate cancer risk: A systematic
review and meta-analysis. Prostate Cancer and Prostatic Diseases, 20(4), 361–377.
https://doi.org/10.1038/pcan.2017.25.
Sabour-Pickett, S., Nolan, J. M., Loughman, J., & Beatty, S. (2012). A review of the
evidence germane to the putative protective role of the macular carotenoids for age-
related macular degeneration. Molecular Nutrition & Food Research, 56(2), 270–286.
https://doi.org/10.1002/mnfr.201100219.
Saini, R. K., & Keum, Y.-S. (2018). Carotenoid extraction methods: A review of recent
developments. Food Chemistry, 240, 90–103. https://doi.org/10.1016/j.foodchem.
2017.07.099.
Saini, R. K., & Keum, Y.-S. (2019). Microbial platforms to produce commercially vital
carotenoids at industrial scale: An updated review of critical issues. Journal of
Industrial Microbiology & Biotechnology, 46(5), 657–674. https://doi.org/10.1007/
s10295-018-2104-7.
Sajilata, M., Singhal, R., & Kamat, M. (2008). The carotenoid pigment zeaxanthin—a
review. Comprehensive Reviews in Food Science and Food Safety, 7(1), 29–49. https://
doi.org/10.1111/j.1541-4337.2007.00028.x.
Sepahi, S., Mohajeri, S. A., Hosseini, S. M., Khodaverdi, E., Shoeibi, N., Namdari, M., &
Tabassi, S. A. S. (2018). Effects of Crocin on diabetic maculopathy: A placebo-con-
trolled randomized clinical trial. American Journal of Ophthalmology, 190, 89–98.
https://doi.org/10.1016/j.ajo.2018.03.007.
Shen, H.-J., Cheng, B.-Y., Zhang, Y.-M., Tang, L., Li, Z., Bu, Y. F., ... Liu, J.-Z. (2016).
Dynamic control of the mevalonate pathway expression for improved zeaxanthin
production in Escherichia coli and comparative proteome analysis. Metabolic en-
gineering, 38, 180–190. https://doi.org/10.1016/j.ymben.2016.07.012.
Shepherd, D., Dasek, J., & Carels, M. S. C. (1976). Production of zeaxanthin. U.S. Patent
No. 3,951,743. Washington, DC: U.S. Patent and Trademark Office.
Shindo, K., & Misawa, N. (2014). New and rare carotenoids isolated from marine bacteria
and their antioxidant activities. Marine Drugs, 12(3), 1690–1698. https://doi.org/10.
3390/md12031690.
Simova, E. D., Frengova, G. I., & Beshkova, D. M. (2003). Effect of aeration on the pro-
duction of carotenoid pigments by Rhodotorula rubra-lactobacillus casei subsp. casei
co-cultures in whey ultrafiltrate. Zeitschrift für Naturforschung C, 58(3–4), 225–229.
https://doi.org/10.1515/znc-2003-3-415.
Su, A., Chi, S., Li, Y., Tan, S., Qiang, S., Chen, Z., & Meng, Y. (2018). Metabolic redesign of
Rhodobacter sphaeroides for lycopene production. Journal of Agricultural and Food
Chemistry, 66(23), 5879–5885. https://doi.org/10.1021/acs.jafc.8b00855.
Sun, T., Miao, L., Li, Q., Dai, G., Lu, F., Liu, T., Zhang, X., & Ma, Y. (2014). Production of
lycopene by metabolically-engineered Escherichia coli. Biotechnology Letter, 36(7),
1515–1522. https://doi.org/10.1007/s10529-014-1543-0.
Suwaleerat, T., Thanapimmetha, A., Srisaiyoot, M., Chisti, Y., & Srinophakun, P. (2018).
Enhanced production of carotenoids and lipids by Rhodococcus opacus PD630. Journal
of Chemical Technology & Biotechnology, 93(8), 2160–2169. https://doi.org/10.1002/
jctb.5554.
Szakács, G., Özvegy, C., Bakos, É., Sarkadi, B., & Váradi, A. (2000). Transition-state
formation in ATPase-negative mutants of human MDR1 protein. Biochemical and
Biophysical Research Communications, 276(3), 1314–1319. https://doi.org/10.1006/
bbrc.2000.3576.
Taniguchi, H., Henke, N. A., Heider, S. A., & Wendisch, V. F. (2017). Overexpression of
the primary sigma factor gene sigA improved carotenoid production by
Corynebacterium glutamicum: Application to production of β-carotene and the non-
native linear C50 carotenoid bisanhydrobacterioruberin. Metabolic Engineering
Communications, 4, 1–11. https://doi.org/10.1016/j.meteno.2017.01.001.
Tian, B., & Hua, Y. (2010). Carotenoid biosynthesis in extremophilic Deinococcus-Thermus
bacteria. Trends in Microbiology, 18(11), 512–520. https://doi.org/10.1016/j.tim.
2010.07.007.
Toyomizu, M., Sato, K., Taroda, H., Kato, T., & Akiba, Y. (2001). Effects of dietary
Spirulina on meat colour in muscle of broiler chickens. British Poultry Science, 42(2),
197–202. https://doi.org/10.1080/00071660120048447.
Üstün, C., Beksac, M., Dalva, K., Koc, H., Konuk, N., Ilhan, O., ... Hayran, M. (2002). In
S. Ram, et al.
vivo use of all-trans retinoic acid prior to induction chemotherapy improves complete
remission rate and increases rhodamine 123 uptake in patients with de novo acute
myeloid leukemia. Medical Oncology, 19(1), 59–67. https://doi.org/10.1385/
MO:19:1:59.
Veiga-Crespo, P., Blasco, L., Rosa dos Santos, F., Poza, M., & Villa, T. G. (2005). Influence
of culture conditions of Gordonia jacobaea MV-26 on canthaxanthin production.
International Microbiology, 8(1), 55–58.
Virtamo, J., Taylor, P. R., Kontto, J., Männistö, S., Utriainen, M., Weinstein, S. J., ...
Albanes, D. (2014). Effects of α-tocopherol and β-carotene supplementation on
cancer incidence and mortality: 18-Year postintervention follow-up of the Alpha-
Tocopherol, Beta-Carotene Cancer Prevention Study. International Journal of Cancer,
135(1), 178–185. https://doi.org/10.1002/ijc.28641.
Viuda-Martos, M., Sanchez-Zapata, E., Sayas-Barberá, E., Sendra, E., Pérez-Álvarez, J., &
Fernández-López, J. (2014). Tomato and tomato byproducts. Human health benefits
of lycopene and its application to meat products: A review. Critical Reviews in Food
Science and Nutrition, 54(8), 1032–1049. https://doi.org/10.1080/10408398.2011.
623799.
Vila, E., Hornero-Méndez, D., Azziz, G., Lareo, C., & Saravia, V. (2019). Carotenoids from
heterotrophic bacteria isolated from Fildes Peninsula, King George Island. Antarctica.
Biotechnology Reports, 21, e00306. https://doi.org/10.1016/j.btre.2019.e00306.
Vine, A. L., Leung, Y. M., & Bertram, J. S. (2005). Transcriptional regulation of connexin
43 expression by retinoids and carotenoids: Similarities and differences. Molecular
Carcinogenesis: Published in cooperation with the University of Texas MD Anderson
Cancer Center, 43(2), 75–85. https://doi.org/10.1002/mc.20080.
Wang, H., Yang, A., Zhang, G., Ma, B., Meng, F., Peng, M., & Wang, H. (2017).
Enhancement of carotenoid and bacteriochlorophyll by high salinity stress in pho-
tosynthetic bacteria. International Biodeterioration & Biodegradation, 121, 91–96.
https://doi.org/10.1016/j.ibiod.2017.03.028.
Wang, Q., Liu, D., Yang, Q., & Wang, P. (2017). Enhancing carotenoid production in
Rhodotorula mucilaginosa KC8 by combining mutation and metabolic engineering.
Annals of Microbiology, 67(6), 425–431. https://doi.org/10.1007/s13213-017-
1274-2.
Xiao, Y.-D., Huang, W.-Y., Li, D.-J., Song, J.-F., Liu, C.-Q., Wei, Q.-Y., ... Yang, Q. M.
(2018). Thermal degradation kinetics of all-trans and cis-carotenoids in a light-in-
duced model system. Food chemistry, 239, 360–368. https://doi.org/10.1016/j.
foodchem.2017.06.107.
Xu, X., Tian, L., Xu, J., Xie, C., Jiang, L., & Huang, H. (2018). Analysis and expression of
the carotenoid biosynthesis genes from Deinococcus wulumuqiensis R12 in engineered
Escherichia coli. AMB Express, 8(1), 94. https://doi.org/10.1186/s13568-018-0624-1.
Yabuzaki, J. (2017). Carotenoids Database: Structures, chemical fingerprints and dis-
tribution among organisms. Database, 2017. https://doi.org/10.1093/database/
bax004.
13
Journal of Functional Foods 67 (2020) 103867
Yamaguchi, M., Igarashi, A., Uchiyama, S., Sugawara, K., Sumida, T., Morita, S., ...
Kajimoto, Y. (2006). Effect of β-crytoxanthin on circulating bone metabolic markers:
Intake of juice (Citrus unshiu) supplemented with β-cryptoxanthin has an effect in
menopausal women. Journal of Health Science, 52(6), 758–768. https://doi.org/10.
1248/jhs.52.758.
Yang, C., Gao, X., Jiang, Y., Sun, B., Gao, F., & Yang, S. (2016). Synergy between me-
thylerythritol phosphate pathway and mevalonate pathway for isoprene production
in Escherichia coli. Metabolic Engineering, 37, 79–91. https://doi.org/10.1016/j.
ymben.2016.05.003.
Yang, D., Park, S. Y., Park, Y. S., Eun, H., & Lee, S. Y. (2020). Metabolic Engineering of
Escherichia coli for Natural Product Biosynthesis. Trends in Biotechnology. https://doi.
org/10.1016/j.tibtech.2019.11.007.
Yang, Y., Liu, B., Du, X., Li, P., Liang, B., Cheng, X., ... Wang, S. (2015). Complete genome
sequence and transcriptomics analyses reveal pigment biosynthesis and regulatory
mechanisms in an industrial strain, Monascus purpureus YY-1. Scientific Reports, 5,
8331. https://doi.org/10.1038/srep08331.
Yolmeh, M., & Khomeiri, M. (2016). Using physical and chemical mutagens for enhanced
carotenoid production from Rhodotorula glutinis (PTCC 5256). Biocatalysis and
Agricultural Biotechnology, 8, 158–166. https://doi.org/10.1016/j.bcab.2016.09.004.
Yolmeh, M., Khomeiri, M., Ghorbani, M., Ghaemi, E., & Ramezanpour, S. S. (2017). High
efficiency pigment production from Micrococcus roseus (PTCC 1411) under ultraviolet
irradiation. Biocatalysis and Agricultural Biotechnology, 9, 156–161. https://doi.org/
10.1016/j.bcab.2016.12.010.
Zhang, S., Tang, G., Russell, R. M., Mayzel, K. A., Stampfer, M. J., Willett, W. C., & Hunter,
D. J. (1997). Measurement of retinoids and carotenoids in breast adipose tissue and a
comparison of concentrations in breast cancer cases and control subjects. The
American Journal of Clinical Nutrition, 66(3), 626–632. https://doi.org/10.1093/ajcn/
66.3.626.
Zhang, J., Sun, Z., Sun, P., Chen, T., & Chen, F. (2014). Microalgal carotenoids: Beneficial
effects and potential in human health. Food & Function, 5(3), 413–425. https://doi.
org/10.1039/C3FO60607D.
Zhang, J., Li, Q. R., Zhang, M. H., You, Y., Wang, Y., & Wang, Y. H. (2019). Enhancement
of carotenoid biosynthesis in Phaffia rhodozyma PR106 under stress conditions.
Bioscience, Biotechnology, and Biochemistry, 83(12), 2375–2385. https://doi.org/10.
1080/09168451.2019.1650633.
Zhou, Q., Zhang, P., & Zhang, G. (2014). Biomass and carotenoid production in photo-
synthetic bacteria wastewater treatment: Effects of light intensity. Bioresource
Technology, 171, 330–335. https://doi.org/10.1016/j.biortech.2014.08.088.
Zigmantas, D., Hiller, R. G., Sundström, V., & Polívka, T. (2002). Carotenoid to chlor-
ophyll energy transfer in the peridinin–chlorophyll-a–protein complex involves an
intramolecular charge transfer state. Proceedings of the National Academy of Sciences,
99(26), 16760–16765. https://doi.org/10.1073/pnas.262537599.