Skip to main content
SpringerLink
Log in

Microbial production of poly-γ-glutamic acid

  • Review
  • Published:
World Journal of Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Poly-γ-glutamic acid (γ-PGA) is a natural, biodegradable and water-soluble biopolymer of glutamic acid. This review is focused on nonrecombinant microbial production of γ-PGA via fermentation processes. In view of its commercial importance, the emphasis is on l-glutamic acid independent producers (i.e. microorganisms that do not require feeding with the relatively expensive amino acid l-glutamic acid to produce γ-PGA), but glutamic acid dependent production is discussed for comparison. Strategies for improving production, reducing costs and using renewable feedstocks are discussed.

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

Source: Kongklom et al. (2015, 2017)

Similar content being viewed by others

References

  • Ashiuchi M (2011) Analytical approaches to poly-γ-glutamate: quantification, molecular size determination, and stereochemistry investigation. J Chromatogr B 879:3096–3101

    Article  CAS  Google Scholar 

  • Ashiuchi M (2013) Microbial production and chemical transformation of poly-γ-glutamate. Microbiol Biotechnol 6:664–674

    CAS  Google Scholar 

  • Ashiuchi M, Kamei T, Baek DH, Shin SY, Sung MH, Soda K, Yagi T, Misono H (2001) Isolation of Bacillus subtilis (chungkookjang), a poly-γ-glutamate producer with high genetic competence. Appl Microbiol Biotechnol 57:764–769

    Article  CAS  Google Scholar 

  • Ashiuchi M, Kamei T, Misono H (2003) Poly-γ-glutamate synthetase of Bacillus subtilis. J Mol Catal B: Enzym 23:101–106

    Article  CAS  Google Scholar 

  • Bajaj I, Singhal R (2009) Enhanced production of poly (γ-glutamic acid) from Bacillus licheniformis NCIM 2324 by using metabolic precursors. Appl Biochem Biotechnol 159:133–141

    Article  CAS  Google Scholar 

  • Bajaj IB, Singhal RS (2011a) Flocculation properties of poly(γ-glutamic acid) produced from Bacillus subtilis isolate. Food Bioproc Technol 4:745–752

    Article  CAS  Google Scholar 

  • Bajaj I, Singhal R (2011b) Poly (glutamic acid)—an emerging biopolymer of commercial interest. Bioresour Technol 102:5551–5561

    Article  CAS  Google Scholar 

  • Birrer GA, Cromwick AM, Gross RA (1994) γ-Poly(glutamic acid) formation by Bacillus licheniformis 9945a: physiological and biochemical studies. Int J Biol Macromol 16:265–275

    Article  CAS  Google Scholar 

  • Buescher JM, Margaritis A (2007) Microbial biosynthesis of polyglutamic acid biopolymer and application in the biopharmaceutical, biomedical and food industries. Crit Rev Biotechnol 27:1–19

    Article  CAS  Google Scholar 

  • Candela T, Fouet A (2006) Poly-gamma-glutamate in bacteria. Mol Microbiol 60:1091–1098

    Article  CAS  Google Scholar 

  • Chen J, Shi F, Zhang B, Zhu F, Cao WF, Xu ZN, Xu GH, Cen PL (2010) Effects of cultivation conditions on the production of γ-PGA with Bacillus subtilis ZJU-7. Appl Biochem Biotechnol 160:370–377

    Article  CAS  Google Scholar 

  • Choi SH, Park JS, Whang KS, Yoon MH, Choi WY (2004) Production of microbial polymer, poly(γ-glutamic acid) by Bacillus subtilis BS 62. Agric Chem Biotechnol 47:60–64

    CAS  Google Scholar 

  • Cromwick AM, Gross RA (1995) Effects of manganese (II) on Bacillus licheniformis ATCC 9945A physiology and γ-poly(glutamic acid) formation. Int J Biol Macromol 17:259–267

    Article  CAS  Google Scholar 

  • Cromwick AM, Birrer GA, Gross RA (1996) Effects of pH and aeration on γ-poly(glutamic acid) formation by Bacillus licheniformis in controlled batch fermentor cultures. Biotechnol Bioeng 50:222–227

    Article  CAS  Google Scholar 

  • da Silva SB, Cantarelli VV, Ayub MAZ (2014) Production and optimization of poly-γ-glutamic acid by Bacillus subtilis BL53 isolated from the Amazonian environment. Bioproc Biosyst Eng 37:469–479

    Article  Google Scholar 

  • de Cesaro A, da Silva SB, Ayub MAZ (2014) Effects of metabolic pathway precursors and polydimethylsiloxane (PDMS) on poly-(gamma)-glutamic acid production by Bacillus subtilis BL53. J Ind Microbiol Biotechnol 41:1375–1382

    Article  Google Scholar 

  • Do JH, Chang HN, Lee SY (2001) Efficient recovery of γ-poly (glutamic acid) from highly viscous culture broth. Biotechnol Bioeng 76:219–223

    Article  CAS  Google Scholar 

  • Du G, Yang G, Qu Y, Chen J, Lun S (2005) Effects of glycerol on the production of poly(γ-glutamic acid) by Bacillus licheniformis. Process Biochem 40:2143–2147

    Article  CAS  Google Scholar 

  • Huang J, Du YM, Xu GH, Zhang HL, Zhu F, Huang L, Xu ZN (2011) High yield and cost-effective production of poly(γ-glutamic acid) with Bacillus subtilis. Eng Life Sci 11:291–297

    Article  CAS  Google Scholar 

  • Jeong JH, Kim JN, Wee YJ, Ryu HW (2010) The statistically optimized production of poly(γ-glutamic acid) by batch fermentation of a newly isolated Bacillus subtilis RKY3. Bioresour Technol 101:4533–4539

    Article  CAS  Google Scholar 

  • Ju WT, Song YS, Jung WJ, Park RD (2014) Enhanced production of poly-γ-glutamic acid by a newly-isolated Bacillus subtilis. Biotechnol Lett 36:2319–2324

    Article  CAS  Google Scholar 

  • Jung DY, Jung S, Yun JS, Kim JN, Wee YJ, Jang HG, Ryu HW (2005) Influences of cultural medium component on the production of poly(γ-glutamic acid) by Bacillus sp. RKY3. Biotechnol Bioproc Eng 10:289–295

    Article  CAS  Google Scholar 

  • Kongklom N, Luo HZ, Shi ZP, Pechyen C, Chisti Y, Sirisansaneeyakul S (2015) Production of poly-γ-glutamic acid by glutamic acid-independent Bacillus licheniformis TISTR 1010 using different feeding strategies. Biochem Eng J 100:67–75

    Article  CAS  Google Scholar 

  • Kongklom N, Shi ZP, Chisti Y, Sirisansaneeyakul S (2017) Enhanced production of poly-γ-glutamic acid by Bacillus licheniformis TISTR 1010 with environmental controls. Appl Biochem Biotechnol 182:990–999

    Article  CAS  Google Scholar 

  • Kreyenschulte D, Krull R, Margaritis A (2014) Recent advances in microbial biopolymer production and purification. Crit Rev Biotechnol 34:1–15

    Article  CAS  Google Scholar 

  • Kubota H, Matsunobu T, Uotani K, Takebe H, Satoh A, Tanaka T, Taniguchi M (1993) Production of poly(γ-glutamic acid) by Bacillus subtilis F-2-01. Biosci Biotech Biochem 57:1212–1213

    Article  CAS  Google Scholar 

  • Kumar R, Pal P (2015) Fermentative production of poly (γ-glutamic acid) from renewable carbon source and downstream purification through a continuous membrane-integrated hybrid process. Bioresour Technol 177:141–148

    Article  CAS  Google Scholar 

  • Lee SY, Chang HN, Thakur NN, Do JH (2006) Method for manufacturing highly-concentrated polyglutamic acid with additional supply of saccharides. United States Patent 6,989,251 B2

    Google Scholar 

  • Lee NR, Lee SM, Cho KS, Jeong SY, Hwang DY, Kim DS, Hong CO, Son HJ (2014) Improved production of poly-γ-glutamic acid by Bacillus subtilis D7 isolated from Doenjang, a Korean traditional fermented food, and its antioxidant activity. Appl Biochem Biotechnol 173:918–932

    Article  CAS  Google Scholar 

  • Li X, Gou XY, Long D, Ji Z, Hu LF, Xu DH, Liu J, Chen ShW (2014) Physiological and metabolic analysis of nitrate reduction on poly-gamma-glutamic acid synthesis in Bacillus licheniformis WX-02. Arch Microbiol 196:791–799

    Article  CAS  Google Scholar 

  • Luo ZT, Guo Y, Liu JD, Qiu H, Zhao MM, Zou W, Li SB (2016) Microbial synthesis of poly-γ-glutamic acid: current progress, challenges, and future perspectives. Biotechnol Biofuels 9:134

    Article  Google Scholar 

  • Mabrouk M, Abou-Zeid DM, Sabra W (2012) Application of Plackett–Burman experimental design to evaluate nutritional requirements for poly (γ-glutamic acid) production in batch fermentation by Bacillus licheniformis A13. Afr J App Microbiol Res 1:6–18

    Google Scholar 

  • Manocha B, Margaritis A (2010) A novel method for the selective recovery and purification of γ-polyglutamic acid from Bacillus licheniformis fermentation broth. Biotechnol Prog 26:734–742

    Article  CAS  Google Scholar 

  • Meissner L, Kauffmann K, Wengeler T, Mitsunaga H, Fukusaki E, Büchs J (2015) Influence of nitrogen source and pH value on undesired poly(γ-glutamic acid) formation of a protease producing Bacillus licheniformis strain. J Ind Microbiol Biotechnol 42:1203–1215

    Article  CAS  Google Scholar 

  • Moraes LP, Alegre RM, Brito PN (2012) Optimisation of poly(γ-glutamic acid) production by Bacillus velezensis NRRL B-23189 in liquid fermentation with molasses as the carbon source without addition of glutamic acid. Int Rev Chem Eng 4:618–623

    Google Scholar 

  • Ogawa Y, Yamaguchi F, Yuasa K, Tahara Y (1997) Efficient production of γ-polyglutamic acid by Bacillus subtilis (natto) in jar fermenters. Biosci Biotechnol Biochem 61:1684–1687

    Article  CAS  Google Scholar 

  • Ogunleye A, Bhat A, Irorere VU, Hill D, Williams C, Radecka I (2015) Poly-γ-glutamic acid: production, properties and applications. Microbiol 161:1–17

    Article  CAS  Google Scholar 

  • Park C, Choi JC, Choi YH, Nakamura H, Shimanouchi K, Horiuchi T, Misono H, Sewaki T, Soda K, Ashiuchi M, Sung MH (2005) Synthesis of super-high-molecular-weight poly-γ-glutamic acid by Bacillus subtilis subsp. chungkookjang. J Mol Catal B Enzym 35:128–133

    Article  CAS  Google Scholar 

  • Peng YY, Jiang B, Zhang T, Mu WM, Miao M, Hu YF (2015) High-level production of poly(γ-glutamic acid) by a newly isolated glutamate-independent strain, Bacillus methylotrophicus. Process Biochem 50:329–335

    Article  CAS  Google Scholar 

  • Richard A, Margaritis A (2001) Poly(glutamic acid) for biomedical applications. Crit Rev Biotechnol 21:219–232

    Article  CAS  Google Scholar 

  • Richard A, Margaritis A (2003) Optimization of cell growth and poly(glutamic acid) production in batch fermentation by Bacillus subtilis. Biotechnol Lett 25:465–468

    Article  CAS  Google Scholar 

  • Shi F, Xu Z, Cen P (2006) Optimization of γ-polyglutamic acid production by Bacillus subtilis ZJU-7 using a surface-response methodology. Biotechnol Bioproc Eng 11:251–257

    Article  CAS  Google Scholar 

  • Shih IL, Van YT (2001) The production of poly-γ-glutamic acid) from microorganisms and its various applications. Bioresour Technol 79:207–225

    Article  CAS  Google Scholar 

  • Shih IL, Van YT, Yeh LC, Lin HG, Chang YN (2001) Production of a biopolymer flocculant from Bacillus licheniformis and its flocculation properties. Bioresour Technol 78:267–272

    Article  CAS  Google Scholar 

  • Shih IL, Wu PJ, Shieh CJ (2005) Microbial production of a poly(γ-glutamic acid) derivative by Bacillus subtilis. Process Biochem 40:2827–2832

    Article  CAS  Google Scholar 

  • Soliman NA, Berekaa MM, Abdel-Fattah YR (2005) Polyglutamic acid (PGA) production by Bacillus sp. SAB-26: application of Plackett–Burman experimental design to evaluate culture requirements. Appl Microbiol Biotechnol 69:259–267

    Article  CAS  Google Scholar 

  • Sung MH, Park C, Kim CJ, Poo HY, Soda K, Ashiuchi M (2005) Natural and edible biopolymer poly-γ-glutamic acid: synthesis, production, and applications. Chem Rec 5:352–366

    Article  CAS  Google Scholar 

  • Tamang JP, Shin DH, Jung SJ, Chae SW (2016) Functional properties of microorganisms in fermented foods. Front Microbiol 7:578

    Google Scholar 

  • Tanaka T, Fujita K, Takenishi S, Taniguchi M (1997) Existence of an optically heterogeneous peptide unit in poly-(γ-glutamic acid) produced by Bacillus subtilis. J Ferment Bioeng 84:361–364

    Article  CAS  Google Scholar 

  • Tork SE, Aly MM, Alakilli SY, Al-Seeni MN (2015) Purification and characterization of gamma poly glutamic acid from newly Bacillus licheniformis NRC20. Int J Biol Macromol 74:382–391

    Article  CAS  Google Scholar 

  • Wei XT, Ji ZX, Chen SW (2010) Isolation of halotolerant Bacillus licheniformis WX-02 and regulatory effects of sodium chloride on yield and molecular sizes of poly-γ-glutamic acid. Appl Biochem Biotechnol 160:1332–1340

    Article  CAS  Google Scholar 

  • Wu Q, Xu H, Xu L, Ouyang PK (2006) Biosynthesis of poly(γ-glutamic acid) in Bacillus subtilis NX-2: regulation of stereochemical composition of poly(γ-glutamic acid). Process Biochem 41:1650–1655

    Article  CAS  Google Scholar 

  • Wu Q, Xu H, Shi NN, Yao J, Li S, Ouyang PK (2008) Improvement of poly(γ-glutamic acid) biosynthesis and redistribution of metabolic flux with the presence of different additives in Bacillus subtilis CGMCC 0833. Appl Microbiol Biotechnol 79:527–535

    Article  CAS  Google Scholar 

  • Wu Q, Xu H, Liang JF, Yao J (2010a) Contribution of glycerol on production of poly(γ-glutamic acid) in Bacillus subtilis NX-2. Appl Biochem Biotechnol 160:386–392

    Article  CAS  Google Scholar 

  • Wu Q, Xu H, Ying HJ, Ouyang PK (2010b) Kinetic analysis and pH-shift control strategy for poly(γ-glutamic acid) production with Bacillus subtilis CGMCC 0833. Biochem Eng J 50:24–28

    Article  Google Scholar 

  • Xu H, Jiang M, Li H, Lu D, Ouyang P (2005) Efficient production of poly(γ-glutamic acid) by newly isolated Bacillus subtilis NX-2. Process Biochem 40:519–523

    Article  CAS  Google Scholar 

  • Xu ZQ, Feng XH, Zhang D, Tang B, Lei P, Liang JF, Xu H (2014) Enhanced poly(γ-glutamic acid) fermentation by Bacillus subtilis NX-2 immobilized in an aerobic plant fibrous-bed bioreactor. Bioresour Technol 155:8–14

    Article  CAS  Google Scholar 

  • Yao J, Xu H, Shi NN, Cao X, Feng XH, Li S, Ouyang PK (2010) Analysis of carbon metabolism and improvement of γ-polyglutamic acid production from Bacillus subtilis NX-2. Appl Biochem Biotechnol 160:2332–2341

    Article  CAS  Google Scholar 

  • Yoon SH, Do JH, Lee SY, Chang HN (2000) Production of poly-γ-glutamic acid by fed-batch culture of Bacillus licheniformis. Biotechnol Lett 22:585–588

    Article  CAS  Google Scholar 

  • Zhang D, Xu ZQ, Xu H, Feng XH, Li S, Cai H, Wei Y, Ouyang PK (2011) Improvement of poly(γ-glutamic acid) biosynthesis and quantitative metabolic flux analysis of a two-stage strategy for agitation speed control in the culture of Bacillus subtilis NX-2. Biotechnol Bioproc Eng 16:1144–1151

    Article  CAS  Google Scholar 

  • Zhang D, Feng XH, Zhou Z, Zhang Y, Xu H (2012a) Economical production of poly(γ-glutamic acid) using untreated cane molasses and monosodium glutamate waste liquor by Bacillus subtilis NX-2. Bioresour Technol 114:583–588

    Article  CAS  Google Scholar 

  • Zhang D, Feng XH, Li S, Chen F, Xu H (2012b) Effects of oxygen vectors on the synthesis and molecular weight of poly(γ-glutamic acid) and the metabolic characterization of Bacillus subtilis NX-2. Process Biochem 47:2103–2109

    Article  CAS  Google Scholar 

  • Zhang HL, Zhu JZ, Zhu XC, Cai J, Zhang AY, Hong YZ, Huang J, Huang L, Xu ZN (2012c) High-level exogenous glutamic acid-independent production of poly-(γ-glutamic acid) with organic acid addition in a new isolated Bacillus subtilis C10. Bioresour Technol 116:241–246

    Article  CAS  Google Scholar 

  • Zhao CF, Zhang YW, Wei XT, Hu ZB, Zhu FY, Xu L, Luo MF, Liu HZ (2013) Production of ultra-high molecular weight poly-γ-glutamic acid with Bacillus licheniformis P-104 and characterization of its flocculation properties. Appl Biochem Biotechnol 170:562–572

    Article  CAS  Google Scholar 

  • Zhu F, Cai J, Wu XT, Huang J, Huang L, Zhu JZ, Zheng Q, Cen PL, Xu ZN (2013) The main byproducts and metabolic flux profiling of γ-PGA-producing strain B. subtilis ZJU-7 under different pH values. J Biotechnol 164:67–74

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biotechnology, Faculty of Agro-Industry, Kasetsart University, Chatuchak, Bangkok, 10900, Thailand

    Sarote Sirisansaneeyakul, Nuttawut Kongklom & Chaniga Chuensangjun

  2. Center for Advanced Studies in Tropical Natural Resources (CASTNAR), National Research University-Kasetsart University (NRU-KU), Kasetsart University, Chatuchak, Bangkok, 10900, Thailand

    Sarote Sirisansaneeyakul, Nuttawut Kongklom & Chaniga Chuensangjun

  3. Department of Chemical and Biological Engineering, NSF Center for Biorenewable Chemicals (CBiRC), Iowa State University, 4118 Biorenewables Research Laboratory, Ames, IA, 50011-1098, USA

    Mingfeng Cao

  4. The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, China

    Zhongping Shi

  5. School of Engineering, Massey University, Palmerston North, New Zealand

    Yusuf Chisti

Authors
  1. Sarote Sirisansaneeyakul
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mingfeng Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nuttawut Kongklom
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chaniga Chuensangjun
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Zhongping Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yusuf Chisti
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sarote Sirisansaneeyakul.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sirisansaneeyakul, S., Cao, M., Kongklom, N. et al. Microbial production of poly-γ-glutamic acid. World J Microbiol Biotechnol 33, 173 (2017). https://doi.org/10.1007/s11274-017-2338-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s11274-017-2338-y

Keywords

Search

Navigation