Purification and Enzymatic Properties of a Difunctional Glycoside Hydrolase from Aspergillus oryzae HML366
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ORIGINAL RESEARCH ARTICLE
Purification and Enzymatic Properties of a Difunctional Glycoside Hydrolase from Aspergillus oryzae HML366 Yongling Qin1,2
•
Yue Fu1,2 • Qiqian Li1,2 • Fengfeng Luo1,2 • Haiyan He1,2
Received: 22 July 2019 / Accepted: 30 May 2020 Ó Association of Microbiologists of India 2020
Abstract In the study, an extracellular enzyme HML CBH1 was purified from the fermentation solution of Aspergillus oryzae HML366, and characterized by biological and molecular analysis. Following the culturing of A. oryzae HML366 under the optimized conditions for enzyme production, an enzyme named HML CBH1 with a molecular weight of 48 kDa was purified using 3000 Da cellulose ultrafiltration column and anion exchange chromatography. The specific activity of the purified enzyme was 9.65 U/mg, and the optimum temperature and pH for the enzyme were 50 and 5.0 °C, respectively. The enzyme was stable at temperatures below 60 °C and pH ranging from 3.0 to 10.0. The partial amino acid sequence of HML CBH1 was analyzed by time-of-flight mass spectrometry, and Mascot and Blast analysis showed that the HML CBH1 sequence was identical to the protein gi:22138643, belonging to the glycoside hydrolase family 7, and had exoglucanase and endoglucanase activity.
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12088-020-00892-5) contains supplementary material, which is available to authorized users. Yongling Qin, Yue Fu and Qiqian Li contributed equally to this work. & Yongling Qin [email protected] & Haiyan He [email protected] 1
College of Chemistry and Biological Engineering, Hechi University, Yizhou 546300, China
2
Guangxi Colleges Universities Key Laboratory of Exploitation and Utilization of Microbial and Botanical Resources, Yizhou 546300, China
Keywords Aspergillus oryzae HML366 Cellulase Purification Time-of-flight mass spectrometry Glycoside hydrolase family 7
Introduction Plant cellulose is the most abundant and cheapest renewable resource on the earth. Through photosynthesis, plants can produce up to 15.5 9 1010 tons of cellulose, of which the total amount of cellulose and hemicellulose is 8.5 9 1010 tons [1, 2]. Cellulose is a structural polysaccharide of plants (including certain fungi and bacteria) and is a major component of cell wall. Cellulose constitutes about 10% of the dry weight of the leaves, more than 50% of the wood, 70% to 80% of the hemp fiber, and 90% to 98% of the cotton [3]. The gross physical structure and morphology of cellulose are well characterized and it is composed of long, unbranched homopolymers of D-glucose units linked by b-1,4-glycosidic bonds to form a linear polymeric chain of over 10,000 glucose residues [4, 5]. Cellulose can be degraded into glucose by enzymatic hydrolysis and saccharification, and finally to ethanol by biotransformation. The resulting product can be further purified by distillation and used as an energy source to replace non-renewable petroleum resources [6]. In nature, cellulases from bacteria and fungi hyd
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