title: Expression of a glucagon-like peptide-1 analogue, as a therapeutic agent for type II diabetes, with enhanced bioactivity and increased N-terminal homogeneity in Pichia pastoris.
abstract: OBJECTIVE
  To improve the bioactivity and increase the N-terminal homogeneity of a glucagon-like peptide-1 (GLP-1) analogue expressed in Pichia pastoris.
  
  
  RESULTS
  The GLP-1 analogue. GGH, consisting of two tandem mutant GLP-1 (GLP-1[A2G]) fused with the N-terminus of human serum albumin (HSA), was expressed in P. pastoris. We also designed and expressed the novel GLP-1 analogue NGGH, which had a His-tag fused with the N-terminus of GGH and an enterokinase (EK) cleavage site at the fusion junction. The His-tag was removed by EK digestion to yield GGH2, which was subsequently compared with GGH expressed in P. pastoris. The purification recovery of GGH2 was 35 % compared with 23 % for GGH. Furthermore, the bioactivity of GGH2 was 605 % higher than GGH, and N-terminal homogeneity was also improved.
  
  
  CONCLUSIONS
  A simple method for the preparation of GGH2 with a cleavable His-tag was developed, and the resultant protein possessed improved bioactivity and N-terminal homogeneity.
authors.affiliation: Laboratory of Drug Design and Molecular Pharmacology, School of Pharmaceutical Sciences, Jiangnan University, Wuxi, 214122, China.|Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Qian
authors.affiliation: Laboratory of Drug Design and Molecular Pharmacology, School of Pharmaceutical Sciences, Jiangnan University, Wuxi, 214122, China.
authors.forename: Chengyuan
authors.initials: C
authors.lastname: Li
authors.affiliation: Laboratory of Drug Design and Molecular Pharmacology, School of Pharmaceutical Sciences, Jiangnan University, Wuxi, 214122, China.
authors.forename: Xiaohai
authors.initials: X
authors.lastname: Gong
authors.affiliation: Laboratory of Drug Design and Molecular Pharmacology, School of Pharmaceutical Sciences, Jiangnan University, Wuxi, 214122, China.
authors.forename: Charles
authors.initials: C
authors.lastname: Ndawula
authors.affiliation: College of Medicine, Huzhou University, Huzhou, Zhejiang, 313000, China. yelongzijing@163.com.
authors.forename: Jin
authors.initials: J
authors.lastname: Qian
authors.affiliation: Laboratory of Drug Design and Molecular Pharmacology, School of Pharmaceutical Sciences, Jiangnan University, Wuxi, 214122, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, China.
authors.forename: Huazhong
authors.initials: H
authors.lastname: Li
authors.affiliation: Laboratory of Drug Design and Molecular Pharmacology, School of Pharmaceutical Sciences, Jiangnan University, Wuxi, 214122, China. jinjian31@163.com.
authors.forename: Jian
authors.initials: J
authors.lastname: Jin
mesh_terms: D003924:Diabetes Mellitus, Type 2; D005285:Fermentation; D052216:Glucagon-Like Peptide 1 / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007004:Hypoglycemic Agents / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D010843:Pichia / Q000235:genetics*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D007004:Hypoglycemic Agents; D011994:Recombinant Proteins; D052216:Glucagon-Like Peptide 1
keywords: Diabetes mellitus; Enterokinase; Glucagon-like peptide-1; His-tag; Homogeneity; Human serum albumin; Pichia pastoris