Skip to main content
SpringerLink
Log in

Evaluation of commercial soy sauce koji strains of Aspergillus oryzae for γ-aminobutyric acid (GABA) production

  • Natural Products - Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

In this study, four selected commercial strains of Aspergillus oryzae were collected from soy sauce koji. These A. oryzae strains designated as NSK, NSZ, NSJ and NST shared similar morphological characteristics with the reference strain (A. oryzae FRR 1675) which confirmed them as A. oryzae species. They were further evaluated for their ability to produce γ-aminobutyric acid (GABA) by cultivating the spore suspension in a broth medium containing 0.4 % (w/v) of glutamic acid as a substrate for GABA production. The results showed that these strains were capable of producing GABA; however, the concentrations differed significantly (P < 0.05) among themselves. Based on the A. oryzae strains, highest GABA concentration was obtained from NSK (194 mg/L) followed by NSZ (63 mg/L), NSJ (51.53 mg/L) and NST (31.66 mg/L). Therefore, A. oryzae NSK was characterized and the sequence was found to be similar to A. oryzae and A. flavus with 99 % similarity. The evolutionary distance (K nuc) between sequences of identical fungal species was calculated and a phylogenetic tree prepared from the K nuc data showed that the isolate belonged to the A. oryzae species. This finding may allow the development of GABA-rich ingredients using A. oryzae NSK as a starter culture for soy sauce production.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Abbreviations

P :

GABA concentration (mg/L)

S :

Substrate concentration (g/L)

S 0 :

Initial substrate concentration (g/L)

t :

Fermentation time (h−1)

X :

Cell concentration (g/L)

X m :

Maximum cell concentration (g/L)

X 0 :

Initial cell concentration (g/L)

Y p/s :

Yield factor for product on cell (mg/g)

Y x/s :

Yield factor for cell on substrate (g/g)

K s :

Monod growth saturation coefficient (g/L)

μ m :

Maximum specific growth rate (h−1)

References

  1. Abe Y, Umemura S, Sugimoto KI, Hirawa N, Kato Y, Yokoyama N, Yokoyama T, Iwai J, Ishii M (1995) Effect of green tea rich in γ-aminobutyric acid on blood pressure of Dahl salt-sensitive rats. Am J Hypertens 8:74–79

    Article  CAS  PubMed  Google Scholar 

  2. Aoki H, Furuya Y, Endo Y, Fujimoto K (2003) Effect of γ-aminobutyric acid-enriched tempeh-like fermented soybean (GABA-tempeh) on the blood pressure of spontaneously hypertensive rats. Biosci Biotechnol Biochem 67:1806–1808

    Article  CAS  PubMed  Google Scholar 

  3. Bueno DJ, Silva JO, Oliver G (2004) Fungal isolation and enumeration in foods. Methods Mol Biol 268:127–131

    PubMed  Google Scholar 

  4. Chaerun SK (2009) Tempeh waste as a natural, economical carbon and nutrient source: ED-XRF and NCS study. HAYATI J Biosci 16:120

    Article  Google Scholar 

  5. Chen CY, Bechtold AG, Tabor J, Bonham AC (2009) Exercise reduces GABA synaptic input onto nucleus tractus solitarii baroreceptor second-order neurons via NK1 receptor internalization in spontaneously hypertensive rats. J Neurosci 29:2754–2761

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Coleman ST, Fang TK, Rovinsky SA, Turano FJ, Moye-Rowley WS (2001) Expression of a glutamate decarboxylase homologue is required for normal oxidative stress tolerance in Saccharomyces cerevisiae. J Biol Chem 276:244–250

    Article  CAS  PubMed  Google Scholar 

  7. D’Andrea G, Granella F, Cataldini M, Verdelli F, Balbi T (2001) GABA and glutamate in migraine. J Headache Pain 2:s57–s60

    Article  PubMed Central  Google Scholar 

  8. de Oliveira V, Felipe L, Wallau GdL, Silva Loreto EL (2009) Isolation of high quality DNA: a protocol combining “rennet” and glass milk. Electron J Biotechnol 12:11–12

    Article  Google Scholar 

  9. Deacon JW (2013) Fungal biology. Blackwell, Malden

    Google Scholar 

  10. El-Enshasy HA (2007) Filamentous fungal cultures—process characteristics, products, and applications. Bioprocess Value Added Prod Renew Resour New Technol Appl 9:225–261

    Article  Google Scholar 

  11. Elbashiti T, Fayyad A, Elkichaoui A (2010) Isolation and identification of Aspergillus oryzae and the production of soy sauce with new aroma. Pak J Nutr 9:1171–1175

    Article  CAS  Google Scholar 

  12. Fazenda ML, Dias JM, Harvey LM, Nordon A, Edrada-Ebel R, LittleJohn D, McNeil B (2013) Towards better understanding of an industrial cell factory: investigating the feasibility of real-time metabolic flux analysis in Pichia pastoris. Microb Cell Fact 12:1

    Article  Google Scholar 

  13. Fazenda ML, Seviour R, McNeil B, Harvey LM (2008) Submerged culture fermentation of ‘‘higher fungi’’: the macrofungi. Adv App Microbiol 63:33–103

    Article  CAS  Google Scholar 

  14. Frost LD (2004) Glucose assays revisited: experimental determination of the glucose concentration in honey. Chem Educ 9:239–241

    CAS  Google Scholar 

  15. Inoue K, Shirai T, Ochiai H, Kasao M, Hayakawa K, Kimura M, Sansawa H (2003) Blood-pressure-lowering effect of a novel fermented milk containing γ-aminobutyric acid (GABA) in mild hypertensives. Eur J Clin Nutr 57:490–495

    Article  CAS  PubMed  Google Scholar 

  16. Jiang D, Ye Y, Hou J (2011) Studies on screening of higher γ-aminobutyric acid-producing Monascus and optimization of fermentative parameters. Eur Food Res Technol 232:541–547

    Article  CAS  Google Scholar 

  17. Kato Y, Kato Y, Furukawa K, Hara S (2002) Cloning and nucleotide sequence of the glutamate decarboxylase-encoding gene gadA from Aspergillus oryzae. Biosci Biotechnol Biochem 66:2600–2605

    Article  CAS  PubMed  Google Scholar 

  18. Kim JY, Lee MY, Ji GE, Lee YS, Hwang KT (2009) Production of γ-aminobutyric acid in black raspberry juice during fermentation by Lactobacillus brevis GABA100. Int J Food Microbiol 130:12–16

    Article  CAS  PubMed  Google Scholar 

  19. Komatsuzaki N, Shima J, Kawamoto S, Momose H, Kimura T (2005) Production of γ-aminobutyric acid (GABA) by Lactobacillus paracasei isolated from traditional fermented foods. Food Microbiol 22:497–504

    Article  CAS  Google Scholar 

  20. Kumar S, Punekar NS (1997) The metabolism of 4-aminobutyrate (GABA) in fungi. Mycol Res 101:403–409

    Article  CAS  Google Scholar 

  21. Kumar S, Punekar NS, SatyaNarayan V, Venkatesh K (2000) Metabolic fate of glutamate and evaluation of flux through the 4-aminobutyrate (GABA) shunt in Aspergillus niger. Biotechnol Bioeng 67:575–584

    Article  CAS  PubMed  Google Scholar 

  22. Leck A (1999) Preparation of lactophenol cotton blue slide mounts. Community Eye Health 12:24

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Lu W, Sun L, Zhang X, Sun J (2010) Properties of glutamate decarboxylase of Aspergillus oryzae and its application for biotransformation of glutamate into gamma-aminobutyric acid. In: Bioinformatics and biomedical engineering (iCBBE), 2010 4th international conference on IEEE, pp 1–5

  24. D’Anjou MC, Daugulis AJ (2001) A rational approach to improving productivity in recombinant Pichia pastoris fermentation. Biotechnol Bioeng 72:1–11

  25. Mombereau C, Kaupmann K, Froestl W, Sansig G, van der Putten H, Cryan JF (2004) Genetic and pharmacological evidence of a role for GABAB receptors in the modulation of anxiety-and antidepressant-like behavior. Neuropsychopharmacol 29:1050–1062

    Article  CAS  Google Scholar 

  26. Oh S-H, Oh C-H (2003) Brown rice extracts with enhanced levels of GABA stimulate immune cells. Food Sci Biotechnol 12:248–252

    CAS  Google Scholar 

  27. Park K-B, Oh S-H (2007) Production of yogurt with enhanced levels of gamma-aminobutyric acid and valuable nutrients using lactic acid bacteria and germinated soybean extract. Bioresour Technol 98:1675–1679

    Article  CAS  PubMed  Google Scholar 

  28. Prakash MS (2010) A simple unstructured mathematical model for ethanol fermentation by mutant Neurospora crassa. Int J Chem Eng Res 2:13–21

    Google Scholar 

  29. Rhyu MR, Kim EY, Kim HY, Ahn BH, Yang CB (2000) Characteristics of the red rice fermented with fungus Monascus. Food Sci Biotechnol 9:21–26

    Google Scholar 

  30. Roohinejad S, Omidizadeh A, Mirhosseini H, Saari N, Mustafa S, Meor Hussin AS, Hamid A, Abd Manap MY (2011) Effect of pre-germination time on amino acid profile and gamma amino butyric acid (GABA) contents in different varieties of Malaysian brown rice. Int J Food Sci Nutr 14:1386–1399

    CAS  Google Scholar 

  31. Rossetti V, Lombard A (1996) Determination of glutamate decarboxylase by high-performance liquid chromatography. J Chromatogr B Biomed Sci Appl 681:63–67

    Article  CAS  Google Scholar 

  32. Samson RA, Hoekstra ES, Frisvad JC (2004) Introduction to food-and airborne fungi. ASM, Washington

  33. Siragusa S, De Angelis M, Di Cagno R, Rizzello C, Coda R, Gobbetti M (2007) Synthesis of γ-aminobutyric acid by lactic acid bacteria isolated from a variety of Italian cheeses. Appl Environ Microbiol 73:7283–7290

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Sooriyamoorthy S, Silva K, Gunawardhane M, Illeperuma C (2004) Isolation and identification of indigenous Aspergillus oryzae for saccharification of rice starch. Trop Agric Res 16:121–127

    Google Scholar 

  35. Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599

    Article  CAS  PubMed  Google Scholar 

  36. Waagepetersen HS, Sonnewald U, Schousboe A (2007) Glutamine, Glutamate, and GABA: metabolic aspects. In: Lajtha A, Oja S, Schousboe A, Saransaari P (eds) Handbook of neurochemistry and molecular neurobiology: amino acids and peptides in the nervous system. Springer, New York, pp 1–21

  37. Wan-Mohtar WAAQI, Ab Kadir S, Saari N (2016) The morphology of Ganoderma lucidum mycelium in a repeated-batch fermentation for exopolysaccharide production. Biotechnol Rep 11:2-11

  38. Wan Mohtar WAAQI, Latif NA, Harvey LM, McNeil B (2016) Production of exopolysaccharide by Ganoderma lucidum in a repeated-batch fermentation. Biocatal Agric Biotechnol 6:91–101

    Google Scholar 

  39. Wang L, Yokoyama K, Takahasi H, Kase N, Hanya Y, Yashiro K, Miyaji M, Nishimura K (2001) Identification of species in Aspergillus section Flavi based on sequencing of the mitochondrial cytochrome b gene. Int J Food Microbiol 71:75–86

    Article  CAS  PubMed  Google Scholar 

  40. Xu D, Pan L, Zhao H, Zhao M, Sun J, Liu D (2011) Breeding and identification of novel koji molds with high activity of acid protease by genome recombination between Aspergillus oryzae and Aspergillus niger. J Ind Microbiol Biotechnol 38:1255–1265

    Article  CAS  PubMed  Google Scholar 

  41. Yang SY, Lu FX, Lu ZX, Bie XM, Jiao Y, Sun LJ, Yu B (2008) Production of gamma-aminobutyric acid by Streptococcus salivarius subsp. thermophilus Y2 under submerged fermentation. Amino Acids 34:473–478

    Article  CAS  PubMed  Google Scholar 

  42. Yang SY, Lu ZX, Lü FX, Bie XM, Sun LJ, Zeng XX (2006) A simple method for rapid screening of bacteria with glutamate decarboxylase activities. J Rapid Methods Aut Microbiol 14:291–298

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We would like to extend our gratitude to the Ministry of Higher Education (MOHE) Malaysia for the financial support awarded to Prof. Dr. Nazamid Saari under the FRGS (Vot No: 5523380).

Author information

Authors and Affiliations

  1. Fermentation Centre, Strathclyde Institute of Pharmacy and Biomedical Sciences, Strathclyde University, 161 Cathedral Street, Glasgow, G4 0RE, Scotland, UK

    Wan Abd Al Qadr Imad Wan-Mohtar

  2. Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

    Safuan Ab Kadir, Abdulkarim Sabo Mohammed & Nazamid Saari

  3. Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Science, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

    Rosfarizan Mohammad

  4. Department of Food Technology, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

    Sarina Abdul Halim Lim

Authors
  1. Safuan Ab Kadir
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wan Abd Al Qadr Imad Wan-Mohtar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rosfarizan Mohammad
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sarina Abdul Halim Lim
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Abdulkarim Sabo Mohammed
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Nazamid Saari
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Nazamid Saari.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ab Kadir, S., Wan-Mohtar, W.A.A.Q.I., Mohammad, R. et al. Evaluation of commercial soy sauce koji strains of Aspergillus oryzae for γ-aminobutyric acid (GABA) production. J Ind Microbiol Biotechnol 43, 1387–1395 (2016). https://doi.org/10.1007/s10295-016-1828-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10295-016-1828-5

Keywords

Search

Navigation