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食品与发酵工业  2017, Vol. 43 Issue (4): 147-    
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高转化人参皂苷菌株筛选鉴定及发酵培养基优化
方磊,陈红,余晓斌
江南大学生物工程学院,工业生物技术教育部重点实验室,江苏无锡,214122
Isolation of high-conversion ginsenoside strain and optimization of fermentation medium
FANG Lei,CHEN Hong,YU Xiao-bin
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摘要 用含七叶苷柠檬酸铁的平板筛选产β-葡萄糖苷酶的菌株;再用对硝基苯酚-β-葡萄糖苷(pNPG)为底物测定β-葡萄糖苷酶酶活,筛选出6株酶活较高的菌株.利用筛选的菌株以人参皂苷Rbl为底物进行发酵转化,通过薄层色谱(thinlayer chromatograph,TLC)和高效液相色谱法(high performance liquid chromatography,HPLC)检测转化产物的种类和含量,确定转化率最高的菌株.该菌株β-葡萄糖苷酶酶活为0.803 U/mL,人参皂苷Rd转化率为97.234 2%.对其进行形态学、18S rDNA基因序列进行分析,经鉴定该菌株是Saccharomycopsis fibuligera.对发酵培养基进行优化,优化结果是:碳源为乳糖,氮源为玉米粉,金属离子为Ca2.最后对诱导剂进行优化,发现诱导剂对转化率没有显著影响,但是芦丁能够诱导人参皂苷Rd向CK的转化.
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Abstract: Strains producing β-glucosidase were isolated by solid medium containing esculin and ferric.Then six strains having high β-glucosidase activity were selected with p-nitropheny l-β-D-glucopyranoside as substrate.Strain having high ginsenoside bioconversion was finally chosen with Rbl as substrate and converted products were determined by TLC and HPLC.β-glucosidase activity of this strain was 0.803 U/mL and ginsenoside Rd conversation rate was 97.234 2%.The selected strain was identified as Saccharomycopsisfibuligera based on the morphology and nucleotide sequence analysis of the 18S rDNA.Fermentation medium was optimized to contain lactose as carbon source and corn flour as nitrogen source,and Ca2+ as metal ion.Finally inducer was optimized to promote the ginsenoside bioconversion.Results indicated that inducer had little impact on the bioconversion but rutin,a kind of inducer,could promote the bioconversion of Rd to CK.The selected strain with high ginsenoside bioconversion had good potential application.
               出版日期:  2017-04-25      发布日期:  2017-12-01      期的出版日期:  2017-04-25
基金资助: 江苏省普通高校研究生科研创新计划项目(SJLX15_0546)
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