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http://dx.doi.org/10.4014/jmb.1405.05069

Efficient Production of ε-Poly-L-Lysine by Streptomyces ahygroscopicus Using One-Stage pH Control Fed-Batch Fermentation Coupled with Nutrient Feeding  

Liu, Sheng-Rong (Guangdong Institute of Microbiology, Ministry-Guangdong Province Jointly State Key Laboratory of Applied Microbiology)
Wu, Qing-Ping (Guangdong Institute of Microbiology, Ministry-Guangdong Province Jointly State Key Laboratory of Applied Microbiology)
Zhang, Ju-Mei (Guangdong Institute of Microbiology, Ministry-Guangdong Province Jointly State Key Laboratory of Applied Microbiology)
Mo, Shu-Ping (Guangdong Institute of Microbiology, Ministry-Guangdong Province Jointly State Key Laboratory of Applied Microbiology)
Publication Information
Journal of Microbiology and Biotechnology / v.25, no.3, 2015 , pp. 358-365 More about this Journal
Abstract
ε-Poly-L-lysine (ε-PL) is a homopolymer of L-lysine molecules connected between the epsilon amino and alpha carboxyl groups. This polymer is currently used as a natural preservative in food. Insufficient biomass is a major problem in ε-PL fermentation. Here, to improve cell growth and ε-PL productivity, various nitrogen-rich nutrients were supplemented into flask cultures after 16 h cultivation, marking the onset of ε-PL biosynthesis. Yeast extract, soybean powder, corn powder, and beef extract significantly improved cell growth. In terms of ε-PL productivity, yeast extract at 0.5% (w/v) gave the maximum yield (2.24 g/l), 115.4% higher than the control (1.04 g/l), followed by soybean powder (1.86 g/l) at 1% (w/v) and corn powder (1.72 g/l) at 1% (w/v). However, supplementation with beef extract inhibited ε-PL production. The optimal time for supplementation for all nutrients examined was at 16 h cultivation. The kinetics of yeast-extract-supplemented cultures showed enhanced cell growth and production duration. Thus, the most commonly used two-stage pH control fed-batch fermentation method was modified by omitting the pH 5.0-controlled period, and coupling the procedure with nutrient feeding in the pH 3.9-controlled phase. Using this process, by continuously feeding 0.5 g/h of yeast extract, soybean powder, or corn powder into cultures in a 30 L fermenter, the final ε-PL titer reached 28.2 g/l, 23.7 g/l, and 21.4 g/l, respectively, 91.8%, 61.2%, and 45.6% higher than that of the control (14.7 g/l). This describes a promising option for the mass production of ε-PL.
Keywords
Streptomyces; ε-poly-L-lysine; one-stage pH control; fed-batch fermentation; nutrient feeding;
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1 Chen XS, Li S, Liao LJ, Ren XD, Li F, Tang L, et al. 2011. Production of ε-poly-L-lysine using a novel two-stage pH control strategy by Streptomyces sp M-Z18 from glycerol. Bioprocess Biosyst. Eng. 34: 561-567.   DOI
2 Chen XS, Ren XD, Zeng X, Zhao FL, Tang L, Zhang HJ, et al. 2013. Enhancement of ε-poly-L-lysine production coupled with precursor L-lysine feeding in glucose-glycerol cofermentation by Streptomyces sp. M-Z18. Bioprocess Biosyst. Eng. 36: 1843-1849.   DOI   ScienceOn
3 EI-Sersy NA, Abdelwahab AE, Abouelkhiir SS, Abou-Zeid DM, Sabry SA. 2012. Antibacterial and anticancer activity of ε-poly-L-lysine (ε-PL) produced by a marine Bacillus subtilis sp. J. Basic Microbiol. 52: 1-10.   DOI   ScienceOn
4 Eom KD, Park SM, Tran HD, Kim MS, Yu RN, Yoo H. 2007. Dendritic α,ε-poly( L-lysine)s as delivery agents for antisense oligonucleotides. Pharm. Res. 24: 1581-1589.   DOI
5 Feng XH, Xu H, Xu XY, Yao J, Yao Z. 2008. Purification and some properties of ε-poly-L-lysine-degrading enzyme from Kitasatospora sp. CCTCC M205012. Process Biochem. 43: 667-672.   DOI   ScienceOn
6 Hiraki J, Ichikawa T, Ninomiya S, Seki H, Uohama K, Seki H, et al. 2003. Use of ADME studies to confirm the safety of ε-polylysine as a preservative in food. Regul. Toxicol. Pharmacol. 37: 328-340.   DOI   ScienceOn
7 Geng WT, Yang C, Gu YY, Liu RH, Guo WB, Wang XM, et al. 2014. Cloning of ε-poly-L-lysine (ε-PL) synthetase gene from a newly isolated ε-PL producing Streptomyces albulus NK660 and its heterologous expression in Streptomyces lividans. Microbial Biotechnol. 7: 155-164.   DOI   ScienceOn
8 Hamano Y, Yoshida T, Kito M, Nakamori S, Nagasawa T, Takagi H. 2006. Biological function of the pld gene product that degrades ε-poly-L-lysine in Streptomyces albulus. Appl. Microbiol. Biotechnol. 72: 173-181.   DOI
9 Sofos JN, Geornaras I. 2005. Activity of ε-polylysine against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes. J. Food Sci. 72: M404-M408.
10 Yamanaka K, Maruyama C, Takagi H, Hamano Y. 2008. ε-Poly-L-lysine dispersity is controlled by a highly unusual nonribosomal peptide synthetase. Nat. Chem. Biol. 4: 766-772.   DOI   ScienceOn
11 Yamanaka K, Kito N, Imokawa Y, Maruyama C, Utagawa T, Hamano Y. 2010. Mechanism of ε-poly-L-lysine production and accumulation revealed by identification and analysis of an ε-poly-L-lysine-degrading enzyme. Appl. Environ. Microbiol. 76: 5669-5675.   DOI   ScienceOn
12 Yoshida T, Nagasawa T. 2003. ε-Poly-L-lysine: microbial production, biodegradation and application potential. Appl. Microbiol. Biotechnol. 62: 21-26.   DOI
13 Zhang Y, Feng XH, Xu H, Yao Z, Ouyang PK. 2010. ε-Poly-L-lysine production by immobilized cells of Kitasatospora sp. MY 5-36 in repeated fed-batch cultures. Bioresour. Technol. 101: 5523-5527.   DOI   ScienceOn
14 Kawai T, Kubota T, Hiraki J, Izumi Y, Yoshikazu I. 2003. Biosynthesis of ε-poly-L-lysine in a cell-free system of Streptomyces albulus. Biochem. Biophys. Res. Commun. 311: 635-640.   DOI   ScienceOn
15 Bankar SB, Singhal RS. 2011. Improved poly-ε-lysine biosynthesis using Streptomyces noursei NRRL 5126 by controlling dissolved oxygen during fermentation. J. Microbiol. Biotechnol. 21: 652-658.
16 Cheng SS, Lu WY, Park SH, Kang DH. 2010. Control of foodborne pathogens on ready-to-eat beef slurry by ε-polylysine. Int. J. Food Microbiol. 141: 236-241.   DOI   ScienceOn
17 Hiraki J, Hatakeyama M, Morita S, Izumi Y. 1998. Improved ε-poly-L-lysine production of an S-(2-aminoethyl)-L-cysteine resistance mutant of Streptomyces albulus. Seibutsu-kogaku Kaishi 76: 487-493.
18 Huang JM, Wu QP, Liu SR, Zhang JM. 2011. Screening of new ε-polylysine producing strain and structure identification of its product. Microbiol. China 38: 871-877.
19 Kahar P, Iwata T, Hiraki J, Park E, Okabe M. 2001. Enhancement of ε-polylysine production by Streptomyces albulus strain 410 using pH control. J. Biosci. Bioeng. 91: 190-194.   DOI   ScienceOn
20 Kito M, Takimoto R, Yoshida T, Nagasawa T. 2002. Purification and characterization of an ε-poly-L-lysine-degrading enzyme from an ε-poly-L-lysine-producing strain of Streptomyces albulus. Arch. Microbiol. 178: 325-330.   DOI
21 Nishikawa M, Ogawa K. 2002. Distribution of microbes producing antimicrobial ε-poly-L-lysine polymers in soil microflora determined by a novel method. Appl. Environ. Microbiol. 68: 3575-3581.   DOI
22 Li S, Li F, Chen XS, Wang L, Xu J, Tang L, Mao ZG. 2011. Genome shuffling enhanced ε-poly-L-lysine production by improving glucose tolerance of Streptomyces graminearus. Appl. Biochem. Biotechnol. 166: 414-423.   DOI   ScienceOn
23 Liu SR, Wu QP, Zhang JM, Mo SP, Yang XJ, Xiao C. 2012. Enhanced ε-poly-L-lysine production from Streptomyces ahygroscopicus by a combination of cell immobilization and in situ adsorption. J. Microbiol. Biotechnol. 22: 1218-1223.   DOI   ScienceOn
24 Saimura M, Takehara M, Mizukami S, Kataoka K, Hirohara H. 2008. Biosynthesis of nearly monodispersed poly(ε-L-lysine) in Streptomyces species. Biotechnol. Lett. 30: 377-385.   DOI
25 Moccia M, Roviello GN, Bucci EM, Pedone C, Saviano M. 2010. Synthesis of a L-lysine-based alternate alpha,epsilon-peptide: a novel linear polycation with nucleic acids-binding ability. Int. J. Pharm. 397: 179-183.   DOI   ScienceOn
26 Ouyang J, Xu H, Li S, Zhu HY, Chen WW, Zhou J, et al. 2006. Production of ε-poly-L-lysine by newly isolated Kitasatospora sp. PL6-3. Biotechnol. J. 1: 1459-1463.   DOI   ScienceOn
27 Pandey AK, Kumar A. 2014. Improved microbial biosynthesis strategies and multifarious applications of the natural biopolymers epsilon-poly-L-lysine. Process Biochem. 49: 496-505.   DOI   ScienceOn
28 Shih IL, Shen MH. 2006. Optimization of cell growth and poly(ε-lysine) production in batch and fed-batch cultures by Streptomyces albulus IFO 14147. Process Biochem. 41: 1644-1649.   DOI   ScienceOn
29 Shima S, Fukuhara Y, Sakai H. 1982. Inactivation of bacteriophages by ε-poly-L-lysine produced by Streptomyces. Agric. Biol. Chem. 46: 1917-1919.   DOI
30 Shih IL, Shen MH, Van YT. 2006. Microbial synthesis of poly(ε-lysine) and its various applications. Bioresour. Technol. 97: 1148-1159.   DOI   ScienceOn
31 Shima S, Sakai H. 1977. Polylysine produced by Streptomyces. Agric. Biol. Chem. 41: 1907-1909.   DOI
32 hima S, Matsuoka H, Iwamoto T, Sakai H. 1984. Antimicrobial action of ε-poly-L-lysine. J. Antibiot. 37: 1449-1455.   DOI