• Article highlight
  • Article tables
  • Article images

Article History

Received : 12-05-2021

Accepted : 19-05-2021

Available online : 26-07-2021



Article Metrics




Downlaod Files

   


Article Access statistics

Viewed: 79

PDF Downloaded: 56

Yadav: Fungal strains for mycological production of citric acid


Introduction

The production of citric acid by fungal strains is influenced by the carbohydrate sources such as molasses, concentration of molasses, incorporation of trace metals and some other compounds, monitoring of parameters to optimize the levels such as hydrogen ion concentration, temperature and incubation period. A probe is necessary for the above mentioned problems which will give the desired improved yield of citric acid.

Successful production of citric acid using potent strain of fungus is dependent on several factors of which the fungus, the culture conditions1 and the raw materials used are most important. The success of a fermentation process depends to a very extent on the use of right type of fungal strain that can produce the desired end product at a minimum cost and in large quantities. The type of fungal strain used will depend on the product. Aspergillus oryzae 2 is a filamentous fungus used to saccharify rice, other grains and potatoes in making of alcoholic beverages. Aspergillus flavus is a fungal pathogen which causes post-harvest disease in cereal grains and legumes. Aspergillus wenti is used to process soybean. Aspergillus foetidus is used to produce koji for shochu- a distilled Japanese alcoholic beverage and it is also utilized for the production of many useful enzymes that serve differing purposes. Aspergillus niger 3, 4, 5 is a fungus and one of the most common species of the genus Aspergillus. It causes a disease called black mold on certain fruits and vegetables such as grapes, onions, peanuts and is a common contaminant of food.

Fungal strains of the Aspergillus niger group of molds have usually given most successful results both in the laboratory and on an industrial basis. Many of these molds produce high yields, imposes fairly uniform biochemical characteristics and they are easily cultivated and produce a negligible quantity of undesirable end products. In the present investigation the following potent fungal strains of some molds have been employed by the author for the mycological production of citric acid using Molasses as feedstock.

Table 1

Fungal strains employed for mycological production of citric acid

1(K)

Aspergillus oryzae

NCIM-647

2 (L)

Aspergillus flavus

NCIM-650

3 (M)

Aspergillus wentii

NCIM-661

4 (N)

Aspergillus foetidus

NCIM-511

5 (O)

Aspergillus niger

NCIM-683

NCIM stands for national Collection of industrial Microbes. The above citric acid producing strains of molds were employed for mycological production of citric acid and the observation are tabulated as in the Table 1 under heading result and discussion.

Experimental Method

The experiment was carried out with the above species of Aspergillus molds. Culture medium was prepared6 as follows:

Table 2

Sucrose:

0.100g

Malt-Extract:

0.075g

Yeast extract:

0.075g

Peptone :

0.075g

Agar-Agar:

1.00g

Distilled Water:

0.100 ml

pH:

1.8

The pH of the culture medium was adjusted to 1.8 by adding requisite amount of KCl-HCl buffer solution. Malt extract is a sweet substances used as a dietary supplement 7 for bacterial culture medium. Peptone mainly acts as a source of nitrogen and also carbon up to some extent. Agar-Agar is a gelatinous substances8 serves as the primary structural support for the algae’s cell wall.

Microorganism grow in a vessel known as culture tubes. About a dozen of clean and steam sterilized dry culture tubes were filled 5 ml of culture medium each and kept in slanting position. A small quantity of Aspergillus was transferred to the freshly prepared culture tubes then kept in an autoclave maintained at 30 0C 8, 9 for 36 hrs. And then placed in refrigerator.

A small quantum of Aspergillus form culture tube was transferred to the freshly prepared inoculum medium 10, 11, 12, 13, 14, 15, 16 in a conical flask and was kept in an incubator for about 36 hours for proper growth of fungus which is finally employed for the inoculation of production medium to get the citric acid on large scale. The composition of the production medium 17 for the mycological production of citric acid by Aspergillus is as follows

Table 3

Molasses:

20%

NH4NO3:

0.60%

KH2PO4:

0.60%

MgSO4.7H2O:

0.60%

pH:

1.8

After inoculation the fermentor flasks were placed in an incubator maintained at a constant temperature at 30 0C and were analyzed after 6, 8 and 10 days of incubation period for the production of citric acid and molasses unfermented. When the citric acid fermentation is over the fermentation broth is kept at 50 0C to avoid contamination. The broth is drained off (filtered) 18 and the mycelium mat is pressed to remove any acid contained in it. The amount of citric acid 19, 20, 21, 22, 23, 24, 25, 26, 27 in the filtrate was measured by titration with 0.1M NaOH against phenolphthalein as an indicator. 28, 29 Total carbohydrates in the fermentation filtrate at various times was determined as glucose by anthrone-sulphuric acid method. 30, 31

Result and Discussion

Table 4

Sl. No

Isolates

Yield of citric acid * in g/100 ml

1(K)

Aspergillus oryzae NCIM-647

5.764

2(L)

Aspergillus flavus NCIM-650

5.983

3(M)

Aspergillus wentii NCIM-661

4.985

4(N)

Aspergillus foetidus NCIM-511

5.236

5(O)

Aspergillus niger NCIM-683

6.886

The data recorded indicates that the fungal strain designated (O) ie Aspergillus niger NCIM-683 has significant yield of citric acid i.e. 6.886 g/100ml in comparison to other isolates. It is thus concluded that all the citric acid producing fungus are not of equal importance though they are morphologically similar to each other. The author therefore has selected Aspergillus niger NCIM-683 for mycological production of citric acid for further study along with different mutagen.32, 33, 34, 35, 36

Source of Funding

None.

Conflict of Interest

None.

Acknowledgement

I express my sincere gratitude to Professor SP Singh Department of Chemistry Magadh University Bodh Gaya for all support and guidance.

References

1 

A R William D J Weber CDC-Guideline for Disinfection and Sterilization in Healthcare Facilities. Prions2010163https://www.cdc.gov/infectioncontrol/pdf/guidelines/disinfection-guidelines-H.pdf

2 

A Rokas The effect of domestication on the fungal proteomeTrends Genet200925260310.1016/j.tig.2008.11.003

3 

R A Samson J Houbraken R C Summerbell B Flannigan J D Mille Common and important species of fungi and actinomycetes in indoor environments2016321511https://pure.knaw.nl/portal/en/publications/common-and-important-species-of-fungi-and-actinomycetes-in-indoor

4 

M L Abarca M R Bragulat G Castellá F J Cabañes Ochratoxin A production by strains of Aspergillus niger var. nigerAppl Environ Microbiol199460726502610.1128/aem.60.7.2650-2652.1994

5 

E Schuster N D Coleman J Frisvad P V Dijck On the safety of Aspergillus niger - a reviewAppl Microbiol Biotechnol20025944263510.1007/s00253-002-1032-6

6 

P Ratnasamy Maintenance of Culture 2nd Edition NCL PuneIndia1977

7 

British pharmaceutical codex. Pharmaceutical Society of Great Britain19074014https://wellcomecollection.org/works/mfze4eb8

8 

L P S Vandenberghe C R Soccol A Pandey J M Lebeault Microbial production of citric acidBraz Arch Biol Technol19994232637610.1590/s1516-89131999000300001

9 

Y Mohsen M Agarwal Effect of Celastrus paniculatus on Learning, Memory and Serum Biochemistry of Aging Albino RatsInt J Chem Chem Eng2013318190

10 

H Zimmermann A Lindgren W Schuhmann L Gorton Anisotropic Orientation of Horseradish Peroxidase by Reconstitution on a Thiol-Modified Gold ElectrodeChem Eur J19939532249

11 

R H Koenemann 1.A Modification Of The Miller-Muntz Method For Colorimetric Determination Of Lactic AcidJ Biol Chem19401351105910.1016/s0021-9258(18)73164-5

12 

L J Rusdell The Nature of Metaphysical ThinkingNature19451563956217810.1038/156217a0

13 

Z F Villano G Langella [Determination of glycoproteins in blood in man by means of paper electrophoresis. III. Results in pathological cases and comparison with blood protein changes]Biol Soc Ital Biol Sper194332634951

14 

C E Horold Studies in the Genus Eremascus: I. The Rediscovery of Eremascus albus Eidam and Some New Observations concerning its Life-history and CytologyAnn Bot19501412748

15 

I D Turyanitsa I D Olekseyuk I I Kozmanko ChemInform Abstract: SYSTEM SB2TE3-SBJ3 UND EIGENSCHAFTEN DER VERBINDUNG SBTEJChemischer Informationsdienst1973114410.1002/chin.197346018

17 

J N Currie The Citric Acid Fermentation Of Aspergillus NigerJ Biol Chem1917311153710.1016/s0021-9258(18)86708-4

18 

F Yokoya Citric Acid ProductionInd Fermentation Ser199241182

19 

H S Grewal K L Kalra Fungal production of citric acidBiotechnol Adv19951322093410.1016/0734-9750(95)00002-8

20 

A Pandey C R Soccol J A Leon P Nigam Production of Organic Acids by Solid-State Fermentation200111326https://pure.ulster.ac.uk/en/publications/solid-state-fermentation-in-biotechnology-fundamentals-and-applic-2

21 

L P S Vandenberghe C R Soccol A Pandey J M Lebeault Review: Microbial production of citric acidBraz. Arch. Biol. Technol199942263

22 

F Yokoya Citric Acid Production. In: Industrial Fermentation SeriesInt J Pharm Pharm Sci19927182

23 

C P Kubicek M Röhr H J Rehm Citric Acid FermentationCrit Rev Biotechnol1985343317310.3109/07388558509150788

24 

W Jianlong W Xianghua Z Ding Production of citric acid from molasses integrated with in-situ product separation by ion-exchange resin adsorptionBioresource Technol2000753231410.1016/s0960-8524(00)00067-5

25 

D T Friesen W C Babcock D J Brose A R Chambers Recovery of citric acid from fermentation beer using supported-liquid membranesJ Membrane Sci19915621274110.1016/s0376-7388(00)80803-0

26 

T Roukas P Kotzekidou Pretreatment of date syrup to increase citric acid productionEnzyme Microbial Technol1997214273610.1016/s0141-0229(97)00041-0

27 

M Legisa M Gradisnik-Grapulin Sudden substrate dilution induces a higher rate of citric acid production by Aspergillus nigerAppl Environ Microbiol19956172732710.1128/aem.61.7.2732-2737.1995

28 

A Suziki S Sarngbin K Krimura S Usami Direct production of citric acid from starch by a 2-deoxyglucose-resistant mutant strain of Aspergillus nigerJ Ferment Bioeng1996814320310.1016/0922-338X(96)80584-8

29 

A Sakurai H Imai T Ejiri K Endoh S Usami Citric acid production by surface culture using Aspergillus niger: Kinetics and simulationJ Fermentation Bioeng199172115910.1016/0922-338x(91)90139-8

30 

H Hamamci Y D Hang Production of citric acid by immobilized dried reactivatedAspergillus nigerBiotechnol Tech1989351410.1007/bf01876221

31 

K H Tan L B Ferguson C Carlton Conversion of cassava starch to biomass, carbohydrates, and acids by Aspergillus nigerJ Appl Biochem1984618090

32 

S Yadav Mycological Production of citric acid by Aspergillus niger exposed to 3-(α-acetonylbenzyl)-4-hyroxycoumarin sodium saltJ Environ Sci Comput Sci Eng Technol201871322810.24214/jecet.A.7.1.05459.

33 

S Yadav Mycological Production of Citric Acid Exposed to 3-AcetamidocoumarinJ Chem Pharm Res201771549

34 

A Rokas The effect of domestication on the fungal proteomeTrends in genetics: TIG 20096063

35 

R A Samson J Houbraken B Flannigan J D Miller Common and important species of fungi and actinomycetes in indoor environments in Microorganisms in Home and Indoor Work Environments200128792

36 

Cyclopedia of India and of eastern and southern Asia, commercial 2187183https://www.biodiversitylibrary.org/item/211446#page/4/mode/1up



jats-html.xsl

© This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.




Wiki in hindi