1株磷脂酶C产生菌株的筛选、鉴定及发酵条件优化

    Screening, Identification, and Optimization of Fermentation Conditions for a Phospholipase C-producing Strain

    • 摘要: 为挖掘可应用于饲料改良及功能性酶制剂生产的新型微生物资源,以山茶油油坊茶饼堆积土样、山茶油粕为分离材料,筛选得到23株能分解磷脂并产生水解透明圈的菌株,从中筛选获得1株磷脂酶C(PLC)高产菌株FL-11。结合菌落形态、生理生化特征与16S rDNA序列同源性分析对菌株进行分类鉴定,结果显示菌株FL-11与Pseudomonas baetica同源性高达99.5%,将其命名为Pseudomonas baetica FL-11。通过单因素和响应面法优化菌株发酵产酶发酵条件,确定最优产酶培养基配方为葡萄糖22.8 g·L−1、玉米浆11.2 g·L−1、20%卵黄液添加量 1%,在此培养条件下该菌株发酵液PLC酶活可达17.45 U·mL−1

       

      Abstract: To explore novel microbial resources applicable to feed improvement and the production of functional enzyme preparations, this study used accumulated soil samples from camellia oil mills and camellia oil meal as isolation materials. Twenty-three strains capable of decomposing phospholipids and producing hydrolytic clear zones were screened, from which strain FL-11, a high-yield producer of phospholipase C(PLC), was selected. The strain was identified through an analysis of colony morphology, physiological and biochemical characteristics, and 16S rDNA sequence homology. The results showed that strain FL-11 shared a 99.5% homology with Pseudomonas baetica and was thus named Pseudomonas baetica FL-11. The components of the fermentation medium for enzyme production were optimized using single-factor experiments combined with response surface methodology. The optimal medium formulation was determined as follows: glucose 22.8 g·L−1, corn steep liquor 11.2 g·L−1, and 20% egg yolk emulsion at an addition level of 1%. Under these culture conditions, the PLC activity of the fermentation broth of this strain reached 17.45 U·mL−1.

       

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