ISSN print edition: 0366-6352 ISSN electronic edition: 1336-9075 Registr. No.: MK SR 9/7 Published monthly

Cloning and expression of two genes coding endo-β-1,4-glucanases from Trichoderma asperellum PQ34 in Pichia pastoris

Nguyen Hoang Loc, Le My Tieu Ngoc, Hoang Tan Quang, Nguyen Duc Huy, and Ngoc Luong

College of Sciences, Hue University, Hue 530000, Vietnam

E-mail: nhloc@hueuni.edu.vn

Abstract: Two genes coding endo-β-1,4-glucanases were cloned from Trichoderma asperellum PQ34 which was isolated from Thua Thien Hue province, Vietnam. The expression of these genes in Pichia pastoris produced two enzymes with molecular masses of approximately 46 kDa (about 42 kDa of enzymes and 4 kDa of signal peptide). The effects of induction time and temperature, inducer concentration, and culture medium on the endo-β-1,4-glucanase activity were investigated. The results showed that the highest total activities of two endo-β-1,4-glucanases were approximately 4.7 × 10^-8 kat (from Glu1-TA gene) and 7.3 × 10^-8 kat (from Glu2-TA gene) occurred after 4 days of induction using 25 mL L^-1 methanol at 30°C when the yeast cells were cultured in a YPL medium.

Keywords: endo-β-1,4-glucanase – expression – Glu1-TA – Glu2-TA – Trichoderma asperellum

Full paper is available at www.springerlink.com.

DOI: 10.1515/chempap-2015-0210

Chemical Papers 70 (3) 284–293 (2016)