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中国精品科技期刊2020
李楠, 游春苹, 刘振民. NADH氧化酶表达对干酪乳杆菌LC2W胞外多糖产量的影响[J]. 食品工业科技, 2016, (06): 182-186. DOI: 10.13386/j.issn1002-0306.2016.06.029
引用本文: 李楠, 游春苹, 刘振民. NADH氧化酶表达对干酪乳杆菌LC2W胞外多糖产量的影响[J]. 食品工业科技, 2016, (06): 182-186. DOI: 10.13386/j.issn1002-0306.2016.06.029
LI Nan, YOU Chun-ping, LIU Zhen-min. Impact of overexpression of NADH oxidase on exopolysaccharide production in Lactobacillus casei LC2W[J]. Science and Technology of Food Industry, 2016, (06): 182-186. DOI: 10.13386/j.issn1002-0306.2016.06.029
Citation: LI Nan, YOU Chun-ping, LIU Zhen-min. Impact of overexpression of NADH oxidase on exopolysaccharide production in Lactobacillus casei LC2W[J]. Science and Technology of Food Industry, 2016, (06): 182-186. DOI: 10.13386/j.issn1002-0306.2016.06.029

NADH氧化酶表达对干酪乳杆菌LC2W胞外多糖产量的影响

Impact of overexpression of NADH oxidase on exopolysaccharide production in Lactobacillus casei LC2W

  • 摘要: 为了考察NADH氧化酶基因(nox)对于干酪乳杆菌LC2W胞外多糖(EPS)产量的影响,本研究以变异链球菌(Streptococcus mutans)基因组为模板,PCR扩增得到1374 bp的nox序列,并构建乳杆菌重组表达质粒p IB184-nox。通过电转化,构建得到一株过表达NADH氧化酶基因的重组干酪乳杆菌LC-nox。该重组菌在无氧发酵时,NADH氧化酶活力为0.7 U/m L,在有氧发酵时,NADH氧化酶活力达到2.65 U/mL。通过HPLC检测发现,发酵液中的乳酸含量随NADH氧化酶活力升高而降低。节省的碳源用于EPS合成,使得重组菌LC-nox在有氧条件下EPS产量最高达263.7 mg/L,比出发菌株提高75.4%。为乳杆菌EPS合成调控提供了新的理论基础。 

     

    Abstract: To investigate the effect of NADH oxidase gene(nox) on the production of exopolysaccharide from Lactobacillus casei LC2 W,nox gene was obtained by PCR amplification with the genomic DNA of Streptococcus mutans as template. It was subcloned into a lactobacilli vector to construct the recombinant expression plasmid p IB184-nox. By electroporation,p IB184-nox was introduced into Lactobacillus casei LC2 W,resulting in the recombinant strain designated as LC-nox. Under anaerobic condition,LC-nox overexpressed NADH oxidase to a level of 0.7 U/m L,while it was elevated to 2.65 U/m L under aerobic condition. Detected by HPLC,the lactate concentration in the fermentation broth showed negative correlation with NADH oxidase activity. The carbon source saved was used for the biosynthesis of exopolysaccharide(EPS). The highest yield of EPS was reached at 263.7 mg/L under aerobic fermentation,which was increased by 75.4% compared to that of the starting strain. A novel theoretical basis was provided for the regulation of EPS biosynthesis in lactobacillus.

     

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