外源氧化胁迫提高透明质酸分子量
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摘要
采用氧化胁迫技术手段,研究了氧化剂过氧化氢对兽疫链球菌Streptococcus zooepidemicus发酵生产透明质酸分子量的影响。经过摇瓶发酵试验表明,外源添加过氧化氢对菌株产生的透明质酸分子量影响显著。在此基础上,进一步考察了5 L发酵罐中添加过氧化氢对透明质酸分子量大小的影响。结果表明:摇瓶水平上,在发酵中期添加过氧化氢有利于透明质酸分子量的提高。发酵罐水平上,发酵7 h时添加过氧化氢溶液,最终透明质酸产量与对照组相比从3.85 g/L提高到4.82 g/L,分子量由1.8×106提高到2.1×106。
The effect of oxidative stress on molecular weight of hyaluranic acid produced by Streptococcus zooepidemicus was investigated.Shake flask experiments suggested that H2O2 had significant effect on molecular weight of hyaluronic acid.The results showed that it was beneficial to enhance the molecular weight of hyaluronic acid when H2O2 was added into the shake flask at the metaphase of fermentation.Further studies were also investigated in the 5 L fermentor system.The results showed that the yield of hyaluronic acid increased from 3.85 g/L to 4.82 g/L,the molecular weight increased from 1.8×106 to 2.1×106 by adding H2O2 in the fermentation time of 7 h.
The effect of oxidative stress on molecular weight of hyaluranic acid produced by Streptococcus zooepidemicus was investigated.Shake flask experiments suggested that H2O2 had significant effect on molecular weight of hyaluronic acid.The results showed that it was beneficial to enhance the molecular weight of hyaluronic acid when H2O2 was added into the shake flask at the metaphase of fermentation.Further studies were also investigated in the 5 L fermentor system.The results showed that the yield of hyaluronic acid increased from 3.85 g/L to 4.82 g/L,the molecular weight increased from 1.8×106 to 2.1×106 by adding H2O2 in the fermentation time of 7 h.
引文
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[2]Zhang X,Duan XJ,Tan WS.Mechanism for the effect of agitation on the molecular weight of hyaluronic acid produced by Streptococcus zooepidemicus[J].Food Chem,2010,119(4):1643-1646.
[3]高海军,陈坚,章燕芳,等.营养条件对兽疫链球菌发酵生产透明质酸的影响[J].生物工程学报,2002,16(3):396-399.
[4]张容鹤,白洋,冯建成,等.链球菌合成透明质酸的研究进展[J].生物技术通报,2010,(4):63-68.
[5]Ogrodowski CS,Hokka CO,Santana MH.Production of hyaluronic acid by Streptococcus:the effects of the addition of lysozyme and aeration on the formation and the rheological properties of the product[J].Appl Biochem Biotechnol,2005,121-124:753-761.
[6]Dukan S,Touati D.Hypochlorous acid stress in Escherichia coli:resistance,DNA damage and comparison with hydrogen peroxide stress[J].J Bacteriol,1996,178(4):6145-6150.
[7]Cleary PP,Larkin A.Hyaluronic acid capsule:strategy for oxygen resistance in group A streptococci[J].J Bacteriol,1979,140(3):1090-1097.
[8]Chong BF,Nielsen LK.Amplifying the cellular reduction potential of Streptococcus zooepidemicus[J].J Biotechnol,2003,100(1):153-162.
[9]Duan XJ,Yang L,Zhang X,et al.Effect of oxygen and shear stress on molecular weight of hyaluronic acid[J].J Microbiol Biotechnol,2008,18(4):718-724.
[10]Blank LM,Hugenholtz P,Nielsen LK.Evolution of the hyaluronic acid synthesis(has)operon in Streptococcus zooepidemicus and other pathogenic streptococci[J].J Mol Evol,2008,67(1):13-22.
[11]Bentley SD,Aanensen DM,Mavroidi A.Genetic analysis of the capsular biosynthetic locus from all90pneumococcal serotypes[J].PLoS Genet,2006,2(3):e31.
[12]Bitter T,Muir HM.A modified uronic acid carbazole reaction[J].Anal Biochem,1962,4(6):330-334.
[13]Rangaswy V,Jain D.An efficient process for production and purification of hyaluronic acid from Streptococcus equi subsp.zooepidemicus[J].Biotechnol Lett,2008,30(3):493-496.
[2]Zhang X,Duan XJ,Tan WS.Mechanism for the effect of agitation on the molecular weight of hyaluronic acid produced by Streptococcus zooepidemicus[J].Food Chem,2010,119(4):1643-1646.
[3]高海军,陈坚,章燕芳,等.营养条件对兽疫链球菌发酵生产透明质酸的影响[J].生物工程学报,2002,16(3):396-399.
[4]张容鹤,白洋,冯建成,等.链球菌合成透明质酸的研究进展[J].生物技术通报,2010,(4):63-68.
[5]Ogrodowski CS,Hokka CO,Santana MH.Production of hyaluronic acid by Streptococcus:the effects of the addition of lysozyme and aeration on the formation and the rheological properties of the product[J].Appl Biochem Biotechnol,2005,121-124:753-761.
[6]Dukan S,Touati D.Hypochlorous acid stress in Escherichia coli:resistance,DNA damage and comparison with hydrogen peroxide stress[J].J Bacteriol,1996,178(4):6145-6150.
[7]Cleary PP,Larkin A.Hyaluronic acid capsule:strategy for oxygen resistance in group A streptococci[J].J Bacteriol,1979,140(3):1090-1097.
[8]Chong BF,Nielsen LK.Amplifying the cellular reduction potential of Streptococcus zooepidemicus[J].J Biotechnol,2003,100(1):153-162.
[9]Duan XJ,Yang L,Zhang X,et al.Effect of oxygen and shear stress on molecular weight of hyaluronic acid[J].J Microbiol Biotechnol,2008,18(4):718-724.
[10]Blank LM,Hugenholtz P,Nielsen LK.Evolution of the hyaluronic acid synthesis(has)operon in Streptococcus zooepidemicus and other pathogenic streptococci[J].J Mol Evol,2008,67(1):13-22.
[11]Bentley SD,Aanensen DM,Mavroidi A.Genetic analysis of the capsular biosynthetic locus from all90pneumococcal serotypes[J].PLoS Genet,2006,2(3):e31.
[12]Bitter T,Muir HM.A modified uronic acid carbazole reaction[J].Anal Biochem,1962,4(6):330-334.
[13]Rangaswy V,Jain D.An efficient process for production and purification of hyaluronic acid from Streptococcus equi subsp.zooepidemicus[J].Biotechnol Lett,2008,30(3):493-496.