控氧发酵对干酪乳杆菌合成苯乳酸的影响
|
摘要
在静置、搅拌及通气搅拌3种不同控氧条件下,分别用干酪乳杆菌Lactobacillus casei B3发酵及全细胞转化合成了苯乳酸,考察菌体生长、葡萄糖消耗及其发酵与转化合成苯乳酸的规律。结果表明:在转速100 r/min的搅拌条件下,L.casei B3发酵合成苯乳酸的浓度比静置发酵条件下提高了41.4%;但在空气流量2 L/min及转速100r/min的通气搅拌下,发酵合成苯乳酸的浓度较静置发酵时下降了60.3%;以8 g/L苯丙酮酸为底物,以相应静置、搅拌及通气搅拌条件下所得的菌体为全细胞催化剂转化合成苯乳酸,其摩尔转化率分别为67.2%、62.7%和35.9%。此结果说明:适度的搅拌促进了发酵过程的底物和产物传质,但充足或过量供氧会影响细胞内的转化合成酶系,不利于苯乳酸的全细胞转化合成。
Phenyl lactic acid was produced by microbial fermentation and whole-cell biotransformation using Lactobacillus casei B3 under different oxygen control conditions,i. e.,the static culture,the agitation,and the aeration-agitation fermentations.Cell growth and glucose consumption were studied. The concentration of phenyl lactic acid was 41. 4% higher in the agitation at the speed of 100 r/min,but60. 3% lower in the aeration-agitation at the speed of 100 r/min and aeration rate of 2 L/min than that in the static culture.Using phenyl pyruvic acid of 8 g/L as substrate,the molar conversion ratios for phenyl lactic acid with the cells from the static culture,the agitation,and the aeration-agitation processes as the whole-cell catalysts,were 67. 2%,62. 7% and 35. 9%,respectively. The results indicated that the mass transfer of substrate and product was enhanced by the suitable agitation,while the activities of key conversion enzymes within the cells could be influenced by the enough oxygen supply regarding the aeration-agitation,and thus was negative to the whole-cell transformation.
Phenyl lactic acid was produced by microbial fermentation and whole-cell biotransformation using Lactobacillus casei B3 under different oxygen control conditions,i. e.,the static culture,the agitation,and the aeration-agitation fermentations.Cell growth and glucose consumption were studied. The concentration of phenyl lactic acid was 41. 4% higher in the agitation at the speed of 100 r/min,but60. 3% lower in the aeration-agitation at the speed of 100 r/min and aeration rate of 2 L/min than that in the static culture.Using phenyl pyruvic acid of 8 g/L as substrate,the molar conversion ratios for phenyl lactic acid with the cells from the static culture,the agitation,and the aeration-agitation processes as the whole-cell catalysts,were 67. 2%,62. 7% and 35. 9%,respectively. The results indicated that the mass transfer of substrate and product was enhanced by the suitable agitation,while the activities of key conversion enzymes within the cells could be influenced by the enough oxygen supply regarding the aeration-agitation,and thus was negative to the whole-cell transformation.
引文
[1]LAVERMICOCCA P,VALERIO F,EVIDENTE A,et al.Purication and characterization of novel antifungal compounds from the sourdough Lactobacillus plantarum strain 21B[J].Appl Environ Microbiol,2000,66(9):4084-4090.
[2]MU W M,CHEN C,LI X F,et al.Optimization of culture medium for the production of phenyllactic acid by Lactobacillus sp.SK007[J].Bioresour Technol,2009,100:1366-1370.
[3]FUJITA T,NGUYEN H D,ITO T,et al.Microbial monomers custom-synthesized to build true bio-derived aromatic polymers[J].Appl Microbiol Biotechnol,2013,97(20):8887-8894.
[4]倪正,关今韬,沈绍传,等.苯乳酸的微生物合成及分离研究进展[J].化工进展,2016,35(11):3627-3633.
[5]ZHENG Z J,ZHAO M Y,ZANG Y,et al.Production of optically pure L-phenyllactic acid by using engineered Escherichia coli coexpressing L-lactate dehydrogenase and formate dehydrogenase[J].J Biotechnol,2015,207:47-51.
[6]朱银龙,贠军贤,沈绍传,等.透化性干酪乳杆菌细胞转化苯丙酮酸合成苯乳酸[J].高校化学工程学报,2015(2):495-500.
[7]王海宽,高雪芹,张淑丽,等.底物对凝结芽孢杆菌TQ33产苯乳酸的影响[J].天津科技大学学报,2014(6):11-15.
[8]KAWAGUCHI H,UEMATSU K,OGINO C,et al.Simultaneous saccharification and fermentation of kraft pulp by recombinant Escherichia coli for phenyllactic acid production[J].Biochem Eng J,2014,88:188-194.
[9]PRASUNA M L,MUJAHID M,SASIKALA C,et al.LPhenylalanine catabolism and L-phenyllactic acid production by a phototrophic bacterium,Rubrivivax benzoatilyticus JA2[J].Microbiol Res,2012,167(9):526-531.
[10]FUJII T,SHIMIZU M,DOI Y,et al.Novel fungal phenylpyruvate reductase belongs to d-isomer-specific 2-hydroxyacid dehydrogenase family[J].Biochim Biophys Acta,2011,1814:1669-1676.
[11]ZHENG Z J,MA C Q,GAO C,et al.Efficient conversion of phenylpyruvic acid to phenyllactic acid by using whole cells of Bacillus coagulans SDM[J].Plo S ONE,2011,6(4):e19030.
[12]韩旭,陈光,王立梅,等.一株产苯乳酸米根霉的鉴定及产物分析[J].吉林农业,2010(4):82-83.
[13]沐万孟,刘凤丽,张涛,等.渗透化细胞转化生成苯乳酸的研究[J].食品与发酵工业,2009,35(3):19-22.
[14]刘凤丽,沐万孟,江波,等.静息细胞转化合成苯乳酸的条件优化[J].食品工业科技,2008,29(8):67-70.
[15]VALERIO F,LAVERMICOCCA P,PASCALE M,et al.Production of phenyllactic acid by lactic acid bacteria:an approach to the selection of strains contributing to food quality and preservation[J].FEMS Microbiol Lett,2004,233:289-295.
[16]THIERRY A,MAILLARD M B.Production of cheese flavor compounds derived from amino acid catabolism by Propionibacterium freudenreichii[J].Dairy Sci Technol,2002,82(1):17-32.
[17]卢志洪,刘志成,郑穗平.不同溶氧条件下谷氨酸棒杆菌发酵过程代谢物及关键酶酶活分析[J].中国酿造,2010,29(6):28-31.
[18]杨艳坤,王芬,孙杨,等.不同溶氧对谷氨酸棒杆菌代谢的影响[J].微生物学通报,2016,43(11):2540-2549.
[19]LI X F,JIANG B,PAN B L,et al.Purification and partial characterization of lactobacillus species SK007 lactate dehydrogenase(LDH)catalyzing phenylpyruvic acid(PPA)conversion into phenyllactic acid(PLA)[J].J Agric Food Chem,2008,56(7):2392-2399.
[2]MU W M,CHEN C,LI X F,et al.Optimization of culture medium for the production of phenyllactic acid by Lactobacillus sp.SK007[J].Bioresour Technol,2009,100:1366-1370.
[3]FUJITA T,NGUYEN H D,ITO T,et al.Microbial monomers custom-synthesized to build true bio-derived aromatic polymers[J].Appl Microbiol Biotechnol,2013,97(20):8887-8894.
[4]倪正,关今韬,沈绍传,等.苯乳酸的微生物合成及分离研究进展[J].化工进展,2016,35(11):3627-3633.
[5]ZHENG Z J,ZHAO M Y,ZANG Y,et al.Production of optically pure L-phenyllactic acid by using engineered Escherichia coli coexpressing L-lactate dehydrogenase and formate dehydrogenase[J].J Biotechnol,2015,207:47-51.
[6]朱银龙,贠军贤,沈绍传,等.透化性干酪乳杆菌细胞转化苯丙酮酸合成苯乳酸[J].高校化学工程学报,2015(2):495-500.
[7]王海宽,高雪芹,张淑丽,等.底物对凝结芽孢杆菌TQ33产苯乳酸的影响[J].天津科技大学学报,2014(6):11-15.
[8]KAWAGUCHI H,UEMATSU K,OGINO C,et al.Simultaneous saccharification and fermentation of kraft pulp by recombinant Escherichia coli for phenyllactic acid production[J].Biochem Eng J,2014,88:188-194.
[9]PRASUNA M L,MUJAHID M,SASIKALA C,et al.LPhenylalanine catabolism and L-phenyllactic acid production by a phototrophic bacterium,Rubrivivax benzoatilyticus JA2[J].Microbiol Res,2012,167(9):526-531.
[10]FUJII T,SHIMIZU M,DOI Y,et al.Novel fungal phenylpyruvate reductase belongs to d-isomer-specific 2-hydroxyacid dehydrogenase family[J].Biochim Biophys Acta,2011,1814:1669-1676.
[11]ZHENG Z J,MA C Q,GAO C,et al.Efficient conversion of phenylpyruvic acid to phenyllactic acid by using whole cells of Bacillus coagulans SDM[J].Plo S ONE,2011,6(4):e19030.
[12]韩旭,陈光,王立梅,等.一株产苯乳酸米根霉的鉴定及产物分析[J].吉林农业,2010(4):82-83.
[13]沐万孟,刘凤丽,张涛,等.渗透化细胞转化生成苯乳酸的研究[J].食品与发酵工业,2009,35(3):19-22.
[14]刘凤丽,沐万孟,江波,等.静息细胞转化合成苯乳酸的条件优化[J].食品工业科技,2008,29(8):67-70.
[15]VALERIO F,LAVERMICOCCA P,PASCALE M,et al.Production of phenyllactic acid by lactic acid bacteria:an approach to the selection of strains contributing to food quality and preservation[J].FEMS Microbiol Lett,2004,233:289-295.
[16]THIERRY A,MAILLARD M B.Production of cheese flavor compounds derived from amino acid catabolism by Propionibacterium freudenreichii[J].Dairy Sci Technol,2002,82(1):17-32.
[17]卢志洪,刘志成,郑穗平.不同溶氧条件下谷氨酸棒杆菌发酵过程代谢物及关键酶酶活分析[J].中国酿造,2010,29(6):28-31.
[18]杨艳坤,王芬,孙杨,等.不同溶氧对谷氨酸棒杆菌代谢的影响[J].微生物学通报,2016,43(11):2540-2549.
[19]LI X F,JIANG B,PAN B L,et al.Purification and partial characterization of lactobacillus species SK007 lactate dehydrogenase(LDH)catalyzing phenylpyruvic acid(PPA)conversion into phenyllactic acid(PLA)[J].J Agric Food Chem,2008,56(7):2392-2399.