摘要
[目的]筛选能高产AMP脱氨酶的野生菌株,并对该菌株进行菌种鉴定和发酵条件优化。[方法]通过大豆的自然发酵制备豆豉曲,并分别以PDA、MRS和YPD 3种培养基进行豆豉曲中微生物的分离纯化,将获得的分离菌株进行液体发酵培养,并检测发酵液的AMP脱氨酶活力。[结果]试验通过3种培养基分离纯化,获得纯种菌株51株。通过进一步的摇瓶发酵培养,检测到具有AMP脱氨酶活性的菌株为16株,选取活性最高的DCP-23菌株进行菌落形态和菌丝形态分析,初步鉴定该菌株为青霉菌属。通过摇瓶发酵条件优化,确定该菌株产酶的适宜发酵条件为:培养基初始pH为6.0,接种量为6%,30℃发酵60 h。在上述发酵条件下,发酵液中的AMP脱氨酶可达到293.7 U/ml。[结论]试验获取了1株能产较高活性AMP脱氨酶的野生青霉菌株DCP-23,可为高产AMP脱氨酶的菌株研究提供参考。
[Objective] To screen out wild strains for producing AMP deaminase,the strain identification and fermentation conditions optimization were conducted.[Method] Through natural fermentation of soybean to prepare Douchiqu,three culture mediums PDA,MRS and YPD were used to conduct separation and purification of microorganism in Douchiqu.The liquid fermentation culture was conducted on the obtained isolated strains,and the AMP deaminase activity was detected.[Result] The strains from Douchiqu were purified respectively with PDA,MRS and YPD medium,and then 51 pure strains in total were obtained.Through further fermentation culture with shake flask,16 strains among them with AMP deaminase activities were detected.The colony morphology and mycelium morphology analysis of DCP-23 with the highest activity were conducted,and this strain was preliminary identified as penicillium.Through optimization of fermentation condition,the suitable condition for enzyme production was as followed：initial pH value of 6.0,inoculation amount of 6％,fermentation of 60 h at 30 ℃.Under the above conditions,the AMP deaminase activity in the fermentation broth could reach 293.7 U/ml.[Conclusion] DCP-23 was obtained,which can provide reference for study on strains producing high yield of AMP deaminase.
出处
《安徽农业科学》
CAS
2014年第27期9532-9536,共5页
Journal of Anhui Agricultural Sciences
基金
江苏省高校自然科学研究面上项目(12KJD550007)
江苏高校优势学科建设工程资助项目
关键词
AMP脱氨酶
自然发酵
豆豉曲
菌株筛选
AMP deaminase
Natural fermentation
Douchiqu
Strain screening
