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嗜热真菌嗜热毁丝霉内切木聚糖酶的生化特性鉴定及其强化生产。

Biochemical characterization and enhanced production of endoxylanase from thermophilic mould Myceliophthora thermophila.

机构信息

Laboratory of Bioprocess Technology, Department of Microbiology, Maharshi Dayanand University, Rohtak, Haryana, 124001, India.

Department of Botany, Pt. N. R. S. Govt. College, Rohtak, Haryana, 124001, India.

出版信息

Bioprocess Biosyst Eng. 2021 Jul;44(7):1539-1555. doi: 10.1007/s00449-021-02539-1. Epub 2021 Mar 25.

Abstract

Endoxylanase production from M. thermophila BJTLRMDU3 using rice straw was enhanced to 2.53-fold after optimization in solid state fermentation (SSF). Endoxylanase was purified to homogeneity employing ammonium sulfate precipitation followed by gel filtration chromatography and had a molecular mass of ~ 25 kDa estimated by SDS-PAGE. Optimal endoxylanase activity was recorded at pH 5.0 and 60 °C. Purified enzyme showed complete tolerance to n-hexane, but activity was slightly inhibited by other organic solvents. Among surfactants, Tweens (20, 60, and 80) and Triton X 100 slightly enhanced the enzyme activity. The V and K values for purified endoxylanase were 6.29 µmol/min/mg protein and 5.4 mg/ml, respectively. Endoxylanase released 79.08 and 42.95% higher reducing sugars and soluble proteins, respectively, which control after 48 h at 60 °C from poultry feed. Synergistic effect of endoxylanase (100 U/g) and phytase (15 U/g) on poultry feed released higher amount of reducing sugars (58.58 mg/feed), soluble proteins (42.48 mg/g feed), and inorganic phosphate (28.34 mg/feed) in contrast to control having 23.55, 16.98, and 10.46 mg/feed of reducing sugars, soluble proteins, and inorganic phosphate, respectively, at 60 °C supplemented with endoxylanase only.

摘要

米黑根毛霉 BJTLRMDU3 在固态发酵(SSF)中经过优化后,内切木聚糖酶的产量提高到 2.53 倍。采用硫酸铵沉淀后凝胶过滤层析对内切木聚糖酶进行纯化,得到电泳纯酶,SDS-PAGE 估计其分子量约为 25 kDa。最佳内切木聚糖酶活性在 pH 5.0 和 60°C 时记录。纯化的酶对正己烷完全耐受,但其他有机溶剂对其活性有轻微抑制。在表面活性剂中,Tween(20、60 和 80)和 Triton X-100 略微提高了酶活性。纯化内切木聚糖酶的 V 和 K 值分别为 6.29µmol/min/mg 蛋白和 5.4mg/ml。与对照相比,在 60°C 下 48 小时后,纯化的内切木聚糖酶分别释放 79.08%和 42.95%更高的还原糖和可溶性蛋白,可控制家禽饲料的质量。与单独添加内切木聚糖酶(100U/g)相比,内切木聚糖酶(100U/g)和植酸酶(15U/g)对家禽饲料的协同作用可释放出更多的还原糖(58.58mg/饲料)、可溶性蛋白(42.48mg/g 饲料)和无机磷(28.34mg/饲料),而对照的还原糖、可溶性蛋白和无机磷分别为 23.55、16.98 和 10.46mg/饲料。

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