The extracellular exopolysaccharide (EPS) produced by Lactobacillus plantarum, a kind of multifunctional lactic acid bacteria, has many excellent functions and properties. However, the high raw materials and operation costs and low EPS yield in scaled fermentations limit its industrial application. The strategies of traditional fed-batch fermentation, bio-catalysis, bio-catalysis combined with liquid-liquid extraction and pH-Stat automatic glucose feeding based “repeated bio-catalysis” were implemented in a 5 L tank. The yield of EPS was 3.34 g/L with glucose by the bio-catalysis method, which was 110% higher than in shake-flask fermentation. With the pH-Stat glucose feeding based repeated bio-catalysis method, the cells harvested in each run could be reused consecutively for the next batch. EPS yield in the second repeated run could still reach a high level of 3.33 g/L. However, the total cell amount gradually decreased due to the cells amount loss during the collection/treatment process, leading to the consecutive decline of EPS concentrations, but the EPS/cell-amount remained constant and the cell metabolic activities were stable. The consecutive reductions of EPS concentration could be solved by enlarging the working volume in the first run and increasing the total cells amounts due to the cells amount loss during the collection/treatment process. The proposed pH-Stat glucose feeding based “repeated bio-catalysis” strategy increased EPS concentration, eliminated the expensive MRS medium utilization, reduced raw materials cost and realized automatic process control. In addition, the proposed strategy could also simultaneously produce 40 g/L DL-lactic acid.
[1] 钟颜麟,彭志英,赵谋明.乳酸菌胞外多糖的研究[J].中国乳品工业,1999,27(4):7-10.
ZHONG Y L,PENG Z Y,ZHAO M M.The studies on exopolysaccharides produced by lactic acid bacteria[J].China Dairy Industry,1999,27(4):7-10.
[2] HUSSAIN A,ZIA K M,TABASUM S,et al.Blends and composites of exopolysaccharides;properties and applications:A review[J].International Journal of Biological Macromolecules,2017,94:10-27.
[3] BENGOA A A,LLAMAS M G,IRAPORDA C,et al.Impact of growth temperature on exopolysaccharide production and probiotic properties of Lactobacillus paracasei strains isolated from kefir grains[J].Food Microbiology,2018,69:212-218.
[4] DEGEEST B,VANINGELGEM F,DE VUYST L.Microbial physiology,fermentation kinetics,and process engineering of heteropolysaccharide production by lactic acid bacteria[J].International Dairy Journal,2001,11(9):747-757.
[5] 胡东良.乳酸菌的抗肿瘤、抗变异原及免疫增强作用[J].中国乳品工业,1997,25(6):9-12.
HU D L.The report on antitumour,antigenic variation and immunological enhancement of lactic acid bacteria[J].China Dairy Industry,1997,25(6):9-12
[6] YASUI H,KAN S D,MATSUZAKI T,et al.Immunomodulatory function of lactic acid bacteria[J].Antonie Van Leeuwenhoek,1999,76(1-4):383-389.
[7] DE VUYST L,DEGEEST B.Heteropolysaccharides from lactic acid bacteria[J].FEMS Microbiology Reviews,1999,23(2):153-177.
[8] 李全阳,夏文水.酸乳中乳酸菌所产胞外多糖特性的初步研究[J].食品科学,2004,25(2):80-84.
LI Q Y,XIA W S.Study on characteristics of exopolysaccharides produced by yoghurt lactic acid bacteria[J].Food Science,2004,25(2):80-84.
[9] 顾笑梅.乳酸菌产胞外多糖的研究[D].济南:山东大学,2003.
GU X M.Studies on excellular polysaccharide produced by lactic acid bacterium[D].Jinan:Shandong University,2003.
[10] 陈东,李程程,史仲平.植物乳杆菌胞外多糖包覆的高稳定性硒纳米颗粒的制备及其抗氧化活性的研究[J].中国生物工程杂志,2020,40(9):18-27.
CHEN D,LI C C,SHI Z P.Lactobacillus plantarum exopolysaccharide coated high-stable selenium nanoparticles and its antioxidant activity[J].China Biotechnology,2020,40(9):18-27.
[11] 王子朝,朱莉,吴剑荣,等.以甘油为底物发酵生产新型黄原胶[J].食品与生物技术学报,2018,37(10):1 028-1 034.
WANG Z C,ZHU L,WU J R,et al.Production of a new xanthan gum with glycerol as substrate[J].Journal of Food Science and Biotechnology,2018,37(10):1 028-1 034.
[12] 王姗杰,詹晓北,吴剑荣,等.低能氮离子注入诱变选育高产结冷胶菌株的研究[J].工业微生物,2010,40(2):20-24.
WANG S J,ZHAN X B,WU J R,et al.Breeding of gellan gum-producing bacteria by low energy N+ ion implantation[J].Industrial Microbiology,2010,40(2):20-24.
[13] 胡立涛,王阳,李佳莲,等.代谢工程改造谷氨酸棒杆菌合成透明质酸[J].食品与发酵工业,2020,46(18):1-7.
HU L T,WANG Y,LI J L,et al.Metabolic engineering of Corynebacterium glutamicum for hyaluronic acid production[J].Food and Fermentation Industries,2020,46(18):1-7.
[14] DE VUYST L,DE VIN F,VANINGELGEM F,et al.Recent developments in the biosynthesis and applications of heteropolysaccharides from lactic acid bacteria[J].International Dairy Journal,2001,11(9):687-707.
[15] MACEDO M G,LACROIX C,GARDNER N J,et al.Effect of medium supplementation on exopolysaccharide production by Lactobacillus rhamnosus RW-9595M in whey permeate[J].International Dairy Journal,2002,12(5):419-426.
[16] MAEDA H,ZHU X,SUZUKI S,et al.Structural characterization and biological activities of an exopolysaccharide kefiran produced by Lactobacillus kefiranofaciens WT-2B(T)[J].Journal of Agricultural and Food Chemistry,2004,52(17):5 533-5 538.
[17] MAEDA H,ZHU X,MITSUOKA T.New medium for the production of exopolysaccharide (OSKC) by Lactobacillus kefiranofaciens[J].Bioscience and Microflora,2003,22(2):45-50.
[18] 陈东.产胞外多糖乳杆菌筛选/鉴定、多糖应用和发酵工艺初步研究[D].无锡:江南大学,2021.
CHEN D.Preliminary study on screening/identification,applications of expolysaccharide and fermentation technology of expolysaccharide-producing Lactobacillus[D].Wuxi:Jiangnan University,2021.
[19] 朱化雷,高大成,孙启梅,等.原位分离技术应用于乳酸发酵的研究进展[J].当代化工,2020,49(9):1 969-1 972;2 032.
ZHU H L,GAO D C,SUN Q M,et al.Research progress in application of in situ separation technology in lactic acid fermentation[J].Contemporary Chemical Industry,2020,49(9):1 969-1972;2 032.
[20] YABANNAVAR V M,WANG D I C.Extractive fermentation for lactic acid production[J].Biotechnology and Bioengineering,1991,37(11):1 095-1 100.
[21] YABANNAVAR V M,WANG D I C.Bioreactor system with solvent extraction for organic acid productiona[J].Annals of the New York Academy of Sciences,1987,506(1):523-535.
[22] BADEL S,BERNARDI T,MICHAUD P.New perspectives for Lactobacilli exopolysaccharides[J].Biotechnology Advances,2011,29(1):54-66.
[23] LI C C,CHEN D,DING J,et al.A novel hetero-exopolysaccharide for the adsorption of methylene blue from aqueous solutions:Isotherm,kinetic,and mechanism studies[J].Journal of Cleaner Production,2020,265:121800.
