为了提高产品中高聚合度ε-聚赖氨酸(ε-poly-L-lysine,ε-PL)的含量,作者在膜筛选基础上,以模拟料液为研究对象,通过单因素实验优化了膜分离工艺参数,比对了分离前后产品的抑菌性能,并分析了不同聚合度ε-PL透过膜的方式。结果表明,截留分子质量为3 000 Da的聚醚砜(polyethersulfone,PES)膜最适合用于筛分聚合度25前后的ε-PL,操作压力为0.25 MPa,pH=6,质量浓度为20 g/L的料液经透析后,循环液中聚合度25~35的ε-PL含量提高至91.22%,透析后的产品更有利于抑制金黄色葡萄球菌和大肠杆菌。膜分离工艺不能实现不同聚合度ε-PL的绝对分离,但是利用不同聚合度ε-PL透过膜的速率不同,可以提高产品中高聚合度ε-PL的比例,从而提高产品的抑菌性能,对于提高ε-PL应用价值具有重要意义。
To increase the content of ε-poly-L-lysine(ε-PL) with high degree of polymerization, membrane was screened and the membrane separation process parameters were optimized. The antibacterial properties of the products before and after separation were compared, while the way ε-PL penetrated the membrane was also analyzed. Results showed that the polyethersulfone(PES) membrane with a molecular weight cut-off of 3 000 Da was the best. At 0.25 MPa, pH 6, and 20 g/L feed solution, the content of ε-PL with a degree of polymerization of 25-35 was increased to 91.22%, and the product after filtering was more conducive to inhibit Staphylococcus aureus and Escherichia coli. The membrane was not able to achieve absolute separation of ε-PL, but it increased the proportion of ε-PL with high polymerization degrees and improved the antibacterial performance. The study is of great significance to improve the application value of ε-PL.
[1] SHIMA S, SAKAI H.Poly-L-lysine produced by Streptomyces.part III.chemical studies[J].Agricultural & Biological Chemistry, 1981, 45(11):2 503-2 508.
[2] SHUKLA S C, SINGH A, PANDEY A K, et al.Review on production and medical applications of ε-poly-lysine[J].Biochemical Engineering Journal, 2012, 65:70-81.
[3] SHIMA S, FUKUHARA Y, SAKAI H.Inactivation of bacteriophages by ε-poly-L-lysine produced by Streptomyces[J].Agricultural and Biological Chemistry, 1982, 46(7):1 917-1 919.
[4] HYLDGAARD M, MYGIND T, VAD B S, et al.The antimicrobial mechanism of action of epsilon-poly-L-lysine[J].Applied and Environmental Microbiology, 2014, 80(24):7 758-7 770.
[5] VAARA M.Agents that increase the permeability of the outer membrane[J].Microbiological Reviews, 1992, 56(3):395-411.
[6] 张东荣, 张超, 段作营, 等.ε-聚赖氨酸抑菌性能的初步研究[J].河南工业大学学报:自然科学版, 2006, 27(3):75-76;80.
ZHANG D R, ZHANG C, DUAN Z Y, et al.A preliminary study on the antibacterial properties of ε-poly-lysine[J].Journal of Henan University of Technology:Natural Science Edition, 2006, 27(3):75-76;80.
[7] PANDEY A K, KUMAR A.Improved microbial biosynthesis strategies and multifarious applications of the natural biopolymer epsilon-poly-L-lysine[J].Process Biochemistry, 2014, 49(3):496-505.
[8] SHUKLA S C, MISHRA A.ε-PolyLysine production from sugar cane molasses by a new isolates of Bacillus sp.and optimization of the fermentation condition[J].Annals of Microbiology, 2013, 63(4):1 513-1 523.
[9] 陈旭升, 李树, 唐蕾, 等.ε-聚赖氨酸生物制造及其在食品防腐领域的应用[J].工业微生物, 2011, 41(4):83-89.
CHEN X S, LI S, TANG L, et al.Bio-manufacturing of ε-poly-lysine and its application in the field of food preservation[J].Industrial Microbiology, 2011, 41(4):83-89.
[10] SHIMA S,MATSUOKA H,IWAMOTO T,et al.Antimicrobial action of ε-poly-L-lysine[J].The Journal of Antibiotics, 1984, 37(11):1 449-1 455.
[11] 刘庆瑞, 陈旭升, 曾昕, 等.链霉菌M-Z18膜蛋白ε-聚赖氨酸降解酶的分离纯化、酶学性质及应用[J].微生物学报, 2014, 54(9):1 022-1 032.
LIU Q R, CHEN X S, ZENG X, et al.Separation, purification, enzymatic properties and application of Streptomyces M-Z18 membrane protein ε-poly-lysine degrading enzyme[J].Acta Microbiologica Sinica, 2014, 54(9):1 022-1 032.
[12] KITO M, TAKIMOTO R, YOSHIDA T,et al.Purification and characterization of an ε-poly-L-lysine-degrading enzyme from an ε-poly-L-lysine-producing strain of Streptomyces albulus[J].Archives of Microbiology, 2002, 178(5):325-330.
[13] NISHIKAWA M, OGAWA K.Inhibition of epsilon-poly-L-lysine biosynthesis in Streptomycetaceae bacteria by short-chain polyols[J].Applied and Environmental Microbiology, 2006, 72(4):2 306-2 312.
[14] 王华, 刘艳飞, 彭东明,等.膜分离技术的研究进展及应用展望[J].应用化工,2013,42(3):532-534.
WANG H, LIU Y F, PENG D M, et al.The development of membrane separation technology and its application prospect[J].Applied Chemical Industry, 2013, 42(3):532-534.
[15] 朱鋆珊, 马平, 郭丽.膜分离技术及其应用[J].当代化工,2017,46(6):1 193-1 195;1 199.
ZHU Y S, MA P, GUO L.Membrane separation technology and its application[J].Contemporary Chemical Industry, 2017, 46(6):1 193-1 195;1 199
[16] ITZHAKI R F.Colorimetric method for estimating poly-lysine and poly-arginine[J].Analytical Biochemistry, 1972, 50(2):569-574.
[17] HIROHARA H, SAIMURA M, TAKEHARA M, et al.Substantially monodispersed poly(-L-lysine)s frequently occurred in newly isolated strains of Streptomyces sp.[J].Applied Microbiology & Biotechnology, 2007, 76(5):1 009-1 016.
[18] XU D L, WANG R, XU Z X, et al.Discovery of a short-chain ε-poly-L-lysine and its highly efficient production via synthetase swap strategy[J].Journal of Agricultural and Food Chemistry, 2019, 67(5):1 453-1 462.
[19] 李芳良, 何洪刚, 马玉, 等.纳滤脱盐在ε-聚赖氨酸分离提取中的应用[J].食品与生物技术学报, 2019, 38(11):122-129.
LI F L, HE H G, MA Y, et al.Application of nanofihration desalination in the separation and extraction of ε-poly-lysine[J].Journal of Food Science and Biotechnology, 2019, 38(11):122-129.
[20] LIU J N, CHANG S L, XU P W, et al.Structural changes and antibacterial activity of epsilon-poly-L-lysine in response to pH and phase transition and their mechanisms[J].Journal of Agricultural and Food Chemistry, 2020, 68(4):1 101-1 109.
