D-pantothenic acid is a kind of vitamin B, which is widely used in feed, food, medicine, and other industries. Due to the high cost and environmental pollution of chemical synthesis of D-pantothenic acid, it is particularly important to use biological fermentation method to produce D-pantothenic acid. In order to achieve high D-pantothenic acid yield by recombinant Escherichia coli DPA21/pBCST3D, some parameters were optimized in the 5 L fermentor, and the feeding mode and concentration of glucose and β-alanine were optimized in the fed-batch fermentation in order to further improve the production of D-pantothenic acid. The optimal conditions were 30 ℃, pH 6.8, and dissolved oxygen (DO) 15%; the pulse feeding of 5-10 g/L glucose was finally selected from two glucose feeding methods, for the subsequent optimization experiment. The optimal β-alanine feeding mode was to mix the β-alanine with glucose solution and the β-alanine concentration was 40 g/L. Under the above fermentation conditions, the titer of D-pantothenic acid reached 41.5 g/L, which was 41.3% higher than that under the initial conditions. The results provide foundations for the industrial production of D-pantothenic acid.
ZHOU Haiyan
,
ZHOU Bin
,
ZOU Shuping
,
ZHANG Bo
,
LIU Zhiqiang
. Optimization of fed-batch fermentation conditions to improve D-pantothenic acid production[J]. Food and Fermentation Industries, 2022
, 48(7)
: 8
-13
.
DOI: 10.13995/j.cnki.11-1802/ts.028317
[1] WEBB M E,SMITH A G,ABELL C.Biosynthesis of pantothenate[J].Natural Product Reports,2004,21(6):695-721.
[2] 杨延辉, 肖春玲.泛酸的功能和生物合成[J].生命的化学,2008,28(4):448-452.
YANG Y H,XIAO C L.The functions and biosynthesis of pantothenate[J].Chemistry of Life,2008,28(4):448-452.
[3] 李明中.D-泛酸钙的应用与合成[J].四川化工与腐蚀控制,1999(2):56-58.
LI M Z.Application and synthesis of calcium D-pantothenate[J].Sichuan Chemical Industry and Corrosion Control,1999(2):56-58.
[4] 孙志浩, 华蕾.生物技术法制备D-泛酸钙和D-泛醇[J].精细与专用化学品,2004,12(10):11-15.
SUN Z H,HUA L.Progress on production of D-calcium pantothenate and D-panthenol by biotechnology[J].Fine and Specialty Chemicals,2004,12(10):11-15.
[5] YAO C X,CHOU J D,WANG T,et al.Pantothenic acid,vitamin C,and biotin play important roles in the growth of Lactobacillus helveticus[J].Frontiers in Microbiology,2018,9:1 194.
[6] EVANS D A,WU J,MASSE C E,et al.A general method for the enantioselective synthesis of pantolactone derivatives[J].Organic Letters,2002,4(20):3 379-3 382.
[7] HEIDLINDEMANN M,HAMMEL M,SCHEFFLER U,et al.Chemoenzymatic synthesis of vitamin B5-intermediate(R)-pantolactone via combined asymmetric organo-and biocatalysis[J].The Journal of Organic Chemistry,2015,80(7):3 387-3 396.
[8] 李伟. 生物催化技术在精细化工中的应用[J].化工设计通讯,2018,44(6):145.
LI W.Application of biocatalysis technology in fine chemical industry[J].Chemical Engineering Design Communications,2018,44(6):145.
[9] 杜明山. 绿色水产饲料添加剂开发现状及前景展望[J].安阳大学学报,2003,2(2):130-132.
DU M S.The present developing situation of green aquatic fodder additives and the future prospect[J].Journal of Anyang University,2003,2(2):130-132.
[10] HIKICHI Y,MORIYA T,MIKI H,et al.Production of D-pantoic acid and D-pantothenic acid:US5518906[P].1996-05-21.
[11] MIYATAKE K,NAKANO Y,KITAOKA S.Pantothenate synthetase of Escherichia coli B I.physicochemical properties[J].The Journal of Biochemistry,1976,79(3):673-678.
[12] MIKI H,YAMAGUCHI T,HIKICHI Y,et al.Production method of D-pantothenic acid and plasmids and microorganisms thereof:Japan,EP19910311927[P].1993-04-14.
[13] RIEPING M,THIERBACH G,PFEFFERLE W,et al.Method for the fermentative production of D-pantothenic acid using strains of the family Enterobacteriaceae:US6184006[P].2001-02-06.
[14] CHASSAGNOLE C,DIANO A,LÉTISSE F,et al.Metabolic network analysis during fed-batch cultivation of Corynebacterium glutamicum for pantothenic acid production:First quantitative data and analysis of by-product formation[J].Journal of Biotechnology,2003,104(1-3):261-272.
[15] SAHM H,EGGELING L.D-pantothenate synthesis in Corynebacterium glutamicum and use of panBC and genes encoding L-valine synthesis for D-pantothenate overproduction[J].Applied and Environmental Microbiology,1999,65(5):1 973-1 979.
[16] ZHANG B,ZHANG X M,WANG W,et al.Metabolic engineering of Escherichia coli for D-pantothenic acid production[J].Food Chemistry,2019,294:267-275.
[17] 于鲁浩, 杨俊慧,孟庆军,等.玉米粉中淀粉含量的快速测定方法[J].山东科学,2012,25(1):19-23.
YU L H,YANG J H,MENG Q J,et al.Fast determination of maize starch content in cornflour[J].Shandong Science,2012,25(1):19-23.
[18] 郑莉莉. D-泛解酸内酯水解酶产生菌的筛选及条件优化的研究[D].济南:山东轻工业学院,2009.
ZHENG L L.Screeming of D-pantolactone hydrolase producing strains and optimizing of technology conditions[D].Jinan:Shandong Polytechnic University,2009.
[19] WANG J Y,RONG H W,ZHANG C S.Evaluation of the impact of dissolved oxygen concentration on biofilm microbial community in sequencing batch biofilm reactor[J].Journal of Bioscience and Bioengineering,2018,125(5):532-542.
[20] 范西强, 陈艳娟,宋敏,等.剪切力对链霉菌谷氨酰胺转胺酶发酵的影响[J].华北农学报,2008,23(S2):126-128.
FAN X Q,CHEN Y J,SONG M,et al.Effects of shear force on microbial transglutaminase fermentation[J].Acta Agriculturae Boreali-Sinica,2008,23(S2):126-128.
[21] 冯志彬, 赵彦宏,王东阳,等.以高浓度葡萄糖为碳源的L-苏氨酸发酵工艺研究[J].中国酿造,2008,27(22):94-96.
FENG Z B,ZHAO Y H,WANG D Y,et al.Study on L-threonine fermentation under high glucose concentration[J].China Brewing,2008,27(22):94-96.
