Rhodobacter sphaeroides

as Microbial Cell Factory

Design, Built, Test, Learn
Concept
Maliha Rashid

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About the Article

Access to Document
10.1002/bit.27593

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Original language English

Impact Factor 4.395

Pages (from-to) 531-541

Journal Biotechnology and Bioengineering

Volume 118

Issue number 2

Early online date 10 Oct 2020

Publication status Published - Feb 2021

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Article Overview

Rhodobacter sphaeroides is a class of bacteria that have the potential to synthesize industrially important products as
microbial cell factories

Learning objectives

• Understanding the microbial cell factories with Rhodobacter sphaeroides as an example

• Understand the DBTL concept
• Analyze how DBTL pipeline is incorporated in Rhodobacter sphaeroides
• Why the use of such a streamlined method for strain engineering is proposed
• Industrial applications of this platform

Keywords

DBTL cycles, industrial biotechnology, metabolic engineering, microbial cell factory,

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What makes R.sphaeroides an ideal choice as a microbial cell
factories?

There are several reasons justifying the interest in R. sphaeroides as chassis for biotechnological productions:

1. Used as a model organism for studying anoxygenic photosynthesis, but also chemotaxis and quorum sensing

2. It displays high metabolic versatility - thrive by aerobic or anaerobic respiration and anoxygenic photosynthesis

3. R. sphaeroides is a natural producer of relevant bio‐ based compounds such as

• Isoprenoids

• poly‐β‐hydroxybutyrate (PHB)

• Hydrogen

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 Overview of the bioeconomy pyramid. This is divided into different sectors based in the market volume
and price per kilo of the compounds produced. In the green boxes, compounds produced by
Rhodobacter sphaeroides are shown. They are allocated to the respective target market sector
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Design, Built, Test, Learn (DBTL) Concept

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Different Research Fields Contributed To The DBTL Method
In R. Sphaeroides

Establishment of a DBTL pipeline in a

chassis requires contribution from
different research fields, including
• ‐omics techniques
• genome engineering
• phenotypic screening methods

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Pathways involved in the carbon metabolism of Rhodobacter sphaeroides

Entner Doudoroff
Pathway

Embden Meyerhoff
Pathway

Mevolonate Pathway
(MVA)

2‐C‐methyl‐D‐erythritol 4‐
phosphate (MEP)
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Functional replacement of isoprenoid pathways in Rhodobacter sphaeroides

Fig: overview of the MEP operon including the target gene dxr. All the other genes
included are involved in growth‐related functions

Fig: overview of the MVA operon from the α‐proteobacterium Paracoccus

zeaxanthinifaciens

Fig: Overview of the strategy

for isoprenoid pathway
replacement in Rhodobacter
sphaeroides.

Microbial Biotechnology, Volume: 13, Issue: 4, Pages: 1082-1093, First published: 24 March 2020, DOI: (10.1111/1751-7915.13562)
Figure: Lumped network of the carbon metabolism of Rhodobacter
sphaeroides, including pathways for substrate uptake and product
formation.

Substrates are highlighted in different colors, each

describing a different growth mode
• light blue: chemoheterotrophic
• light orange: photoheterotrophic
• light yellow: photo‐ or chemolitho‐autotrophic

The three carbon products described in this review

are highlighted
1. 5‐aminolevulinic acid (5‐ALA, yellow)
2. isoprenoids (red)
3. poly‐β‐hydroxybutyrate (PHB, green)

Some pathways with parallel flux are highlighted,

for example, glycolysis:
• Emden–Meyerhof–Parnas (red)
• Entner–Doudoroff (gray)
• isoprenoid synthesis: 2‐C‐methyl‐D‐erythritol 4‐
phosphate (MEP) pathway (blue)
• mevalonate pathway (orange)

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An Outlook on the Industrial Applications Of R. Sphaeroides:
Current Situation and Future Perspectives

This microorganism is considered nonpathogenic and generally regarded as safe (GRAS).

R. sphaeroides is being employed in few companies worldwide:

Company Name Website link Area of bacterial use

produces sesquiterpenes as aroma

Dutch flavors and fragrances company Isobionics BV www.isobionics.com
compunds

Chinese company CN Lab Nutrition www.cnlabnutrition.com Nutraceuticals, coenzyme Q10 with a rate
of up to 30 tons per month.
Chinese company Hebei Shixiang Biological Technology Co.,
www.hbshixiang.en.china.cn feed industry for livestock
Ltd.

Indian company Prions Biotech www.prionsbiotech.com Fish feed solutions

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Future Perspectives
This microorganism could be exploited for evaluating
alternative feedstocks for the bio‐based economy
because of
• its versatile metabolism
• its wide substrate acceptance range
• recent advancements in its DBTL method
the use of cheap and renewable
feedstocks could improve both
commercial and experimental processes
where R. sphaeroides is involved
Technologies used in DBTL pipeline could be
used in R. sphaeroides to optimize this
platform for assimilation of C1 substrates,
while coupling it to synthesis to a range of
bio‐based compounds
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Take home Message

• R. sphaeroides has the potential to produce industrially important bioproducts

• Moreover, it is reasoned that recent improvements in the DBTL pipeline can

be employed for further optimizing this cell factory for industrial applications

of other organisms

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 References

1. Microbial Biotechnology, Volume: 13, Issue: 4, Pages: 1082-1093, First published: 24 March 2020, DOI:
(10.1111/1751-7915.13562)

2. Enrico Orsi, Jules Beekwilder, Gerrit Eggink, Servé W. M. Kengen, Ruud A. Weusthuis. The transition
of Rhodobacter sphaeroides into a microbial cell factory (2020). https://doi.org/10.1002/bit.27593

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min

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