Dr. K.

USHASRI
M.Sc,Ph,D
Govt. Degree College (A), Chittoor
Email. Id : uva1368@gmail.com 1
CONTENTS
 Introduction
 Raw materials and microorganisms
 Biochemical pathway
 Steps involved in Production processes
 Fermentation methods
 Applications of citric acid
 Conclusion
 References 2
LEARNING OBJECTIVES

 Structure of citric acid & bio chemistry of citric acid

 Agricultural waste materials as substrates

 Microbes used for fermentation

 Types of Citric acid fermentation processes.

 Applications of citric acid

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INTRODUCTION

Production of Citric Acid By Supriya N biology reader.com 4

 HISTORICAL DEVELOPMENTS
 1784: Carl Wilhelm Scheele -citric acid was first isolated from the
lemon juice and crystalized.
 1893:Wehmer - Penicilium mold could produce Citric acid

 1917: James Currie - Aspergillus niger could be an efficient citric

acid producer
 1922: Millard observed the accumulation of citric acid in the
Aspergillus.
 1965: yeasts were used for the citric acid production.

 1984: Abould Zeid and Ashy - “Submerged fermentation 5

technique”
 BIOLOGICAL METHOD
About 99% of world production of citric acid occurs via microbial
processes.
The industrial citric acid production can be carried in three
different ways

Surface Process

Citric acid Submerged

fermentation Process

Solid state Process 6

 CITRIC ACID PRODUCERS
Fungi A.niger A.wenti, A. foetidus, Few mutants
A. aculeatus, A. awamori, of A.niger and A. wentii,
A.fumaricus, A. Flavus used for industrial
Trichoderma viride and Mucor pyriformis production.
Ustulina vulgaris,
Pencillium restrictum
Yeast Candida lipolytica, Torulapsis sp ,Rhodotorila sp, Saccharomycopsis sp.
Trichosporon, Saccharomyces are employed for
sp and Zygosaccharomyces sp commercial production.
Bacteria Bacillus Licheniformis, Arthrobacter paraffinens,
Coryne bacterium sps
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 SUBSTRATES
Fermentation
Raw materials
Surface Brewery waste, cotton waste, turnip whey (enriched with molasses)

Sub Bagasse, beet molasses, black strap molasses, brewery wastes, cane
merged molasses, corn starch, date syrup, glycerol, hydrolysate starch, n-
paraffin, palm oil, olive oil, soya bean oil, rapeseed oil, spent grain
liquor, wood hemicelluloses, xylan hydrolysate.
Solid state Wheat bran, carrot waste, cassava bagasse, cellulose hydrolysate
and sugar cane, coffee husk, corncob, rice bran, grape pomace, kiwi
fruit peel, kumara, molasses, okara, orange waste, pineapple waste,
rice bran, sucrose, sugarcane-press mud, wheat bran
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 FERMENTATION STEPS
 The fermentation processes have four phases,irrespective
of the type of fermentation.

 Medium preparation
 Inoculum preparation and addition

 Fermentation

 Recovery of the Citric acid

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 MEDIUM PREPARATION
 Saccharine sources: Sugar, Sugar cane syrup, Sugar cane
molasses,Sugar beet molasses.
 Starchy sources: Potato starch, Starch hydrolysates

 Nitrogen and phosphate sources:

Nitrogen: Na NO3,KNO3 or NH4NO3

Phosphate:K2HPO4 or KH2PO4.
Trace elements: Cu,Mn,Mg,Fe,Zn,Mo in ppm 10
INOCULUM PREPARATION
 Spores------Spore suspension

 Surface process- spores+ Fermentation medium

 Submerged process-Spores + mycelial pellets

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 FERMENTATION CONDITIONS

Surface Submerged Solid

Aeration-Surface Aeration- from Aeration –

diffusion bottom through bran
Temp-25 °C to 30°C Temp- >30 °C Temp- >30 ° C
pH-4-5 pH-3.5 pH-3.0 12
Incubation-7-12 Incubation-4-5 Incubation-5-8
 SURFACE PROCESS

Beet molasses(15-20
% of sucrose, added
nutrients)Inorganic
salts:(NH4)2NO3 ,
MgSO4
28-30 °C, pH 4.0 - 5 .0, 8 12Days
Citric
acid

ferrocyanide or
ferricyanide
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Conidial suspension
 SUBMERGED FERMENTATION
Beet molasses substrate (12
- 15 %.), ammonium nitrate
or potassium dihydrogen
phosphate
>30 ºc,pH3.5 , 5-14Days
Citric
acid
Takes 9 to 25 hours at
32 ºC Mycelia pellets
A.Niger spores are
5 to 25 x 106 per
litre of substrate 14
 SOLID-STATE FERMENTATION

Wheat bran
is moistened Hot water
to about 70% extraction
moisture /Mechanical
>30ºc ,PH-3 , for 5-8Days
press

Aspergillus niger Citric acid

aerosol/liquid
conidia.
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HARVEST AND RECOVERY

Production of Citric Acid By Supriya N biology reader.com 16

 FORMATION OF CITRIC ACID
 The citric acid produces at the time
of trophophase cell growth as a result of
defective citric acid or Kreb’s cycle.

 Defective Kreb’s cycle

 The Kreb’s cycle must be deactivated like an

enzyme Aconitase/ Isocitratedehydrogenase
which can further break down the citric acid.
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biology reader.com
APPLICATIONS OF CITRIC ACID
Food industry

Beverages

Pharmaceuticals

Other industries 18
 Dairy
Candies products
Cosmetics
Fats and oils

Pharmaceuticals
Citric Frozen fruits

acid
Soft drinks and
syrups Jellies and
jams

Wines and Fruit and

ciders vegetable 19
juices
CONCLUSION
 Citric acid is an intermediately prodcut of kerbs' cycle.
 Agricultural waste materials can be used for citric acid
production as ecofriendly process.
 Aspergillus species are widely used for fermentation

 Different fermentation processes can be used for

production
 Citric acid has versatile applications in many areas.

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 REFERENCES
 Text book of microbiology Applied -Telugu akademi,Hyderabad
 Text book of Industrial microbiologyPatel AH. (1996). Industrial
Microbiology .1st Edition.
 Aghdam M.G., Taherzadeh M. Production of citric acid by solid state
fermentation. JBiotechnol. 2008;136:S464.
 Kubicek, C. P. and Rohr, M., (1986) Citric acid fermentation, CRC Crit. Rev.
Biotechnol., 3, 331-373 Aboud-Zeid, A., and Ashy, M. A. (1984) Production
of citric acid: A review. Agric. Wastes, 9, 51-76

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 Overview of citric acid production from Aspergillus niger Pages 271-283 |
Received 19 Oct 2014, Accepted 21 Mar 2015, Published online: 20 Apr
2015
 www.scielo.br
 Pau Loke Show,Kehinde Opeyemi Oladele,Qi Yan Siew,Fitri Abdul Aziz
Zakry,John Chi-Wei Lan &Tau Chuan Ling.
 Currie, J.N. (1917). The citric acid fermentation of A. niger. J. Biol.
Chem. 31, 5.
 CITRIC ACID PRODUCTION Presented by: N.Nachal (18FET210)
Nishank Waghmare (18FET211) Biotechnological production of citric acid
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 Production of Citric Acid By Supriya N ,biology reader.com
 http://www.myandegroup.com/citric-acid-process.
 http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-
89131999000300001
 http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-
83822010000400005
 https://www.slideshare.net/DineshS50/citric-acid-production-74641179
 https://www.sciencedirect.com/science/article/pii/S1387265607130118
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THANK YOU

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