Cloning, expression, biochemical characterization, and molecular docking studies of a novel glucose tolerant β-glucosidase from Saccharomonospora sp. NB11

被引:13
|
作者
Zada, Numan Saleh [1 ,2 ]
Belduz, Ali Osman [2 ]
Guler, Halil Ibrahim [2 ]
Khan, Anum [1 ]
Sahinkaya, Miray [2 ]
Kaciran, Arife [2 ]
Ay, Hilal [3 ]
Badshah, Malik [1 ]
Shah, Aamer Ali [1 ]
Khan, Samiullah [1 ]
机构
[1] Quaid I Azam Univ, Dept Microbiol, Islamabad 45320, Pakistan
[2] Karadeniz Tech Univ, Fac Sci, Dept Mol Biol, TR-61080 Trabzon, Turkey
[3] Ondokuz Mayis Univ, Fac Sci & Arts, Dept Mol Biol & Genet, Samsun, Turkey
关键词
Novel beta-glucosidase; Glucose tolerant; Saccharomonospora sp. NB11; Molecular docking; TRICHODERMA-REESEI; PURIFICATION; HYDROLYSIS; OVEREXPRESSION; INHIBITION; METAGENOME; CELLOBIOSE; ETHANOL;
D O I
10.1016/j.enzmictec.2021.109799
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Most of the presently known beta-glucosidases are sensitive to end-product inhibition by glucose, restricting their potential use in many industrial applications. Identification of novel glucose tolerant beta-glucosidase can prove a pivotal solution to eliminate end-product inhibition and enhance the overall lignocellulosic saccharification process. In this study, a novel gene encoding beta-glucosidase BglNB11 of 1405bp was identified in the genome of Saccharomonospora sp. NB11 and was successfully cloned and heterologously expressed in E. coli BL21 (DE3).The presence of conserved amino acids; NEPW and TENG indicated that BglNB11 belonged to GH1 beta-glucosidases. The recombinant enzyme was purified using a Ni-NTA column, with the molecular mass of 51 kDa, using SDSPAGE analysis. BglNB11 showed optimum activity at 40 degrees C and pH 7 and did not require any tested co-factors for activation. The kinetic values, K-m, V-max, k(cat), and k(cat)/K-m of purified enzyme were 0.4037 mM, 5735.8 mu mol/min/mg, 5042.16 s(-1) and 12487.71 s(-1) mM(-1), respectively. The enzyme was not inhibited by glucose to a concentration of 4 M but was slightly stimulated in the presence of glucose. Molecular docking of BglNB11 with glucose suggested that the relative binding position of glucose in the active site channel might be responsible for modulating end product tolerance and stimulation. beta-glucosidase from BglNB11 is an excellent enzyme with high catalytic efficiency and enhanced glucose tolerance compared to many known glucose tolerant beta-glucosidases. These unique properties of BglNB11 make it a prime candidate to be utilized in many biotechnological applications.
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页数:14
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