title: Expression, purification, and lipolytic activity of recombinant human serum albumin fusion proteins with one domain of human growth hormone in Pichia pastoris.
abstract: Human growth hormone (hGH) can mobilize lipid and inhibit the synthesis of triglycerides. However, it is not a potentially useful drug for treating obesity because it has many other actions resulting in several side effects. Here, we report a novel approach to develop the lipolytic function of hGH. The amino terminus of hGH was replaced by an inactive protein so that the actions unrelated to lipolytic function would be avoided. The fusion genes encoding human serum albumin (HSA) and lipolytic domain of hGH were constructed and expressed in Pichia pastoris. The recombinant proteins were purified and characterized by SDS-PAGE and Western blot. The preliminary stability tests demonstrated that HSA-hGH166-191 and HSA-hGH177-191 were stable at different pH levels after four days at 37°C. Lipolytic activity assay revealed that fusion proteins could increase the amounts of glycerol released from the isolated adipocytes. The HSA fusion proteins constructed in this work can be further developed as antiobesity agents.
authors.affiliation: Institute of Pharmacology & Toxicology and Biochemical Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, People's Republic of China; Ningbo Institute of Medical Science, Medical School, Ningbo University, Ningbo, People's Republic of China.
authors.forename: Furong
authors.initials: F
authors.lastname: Wang
authors.forename: Min
authors.initials: M
authors.lastname: Wu
authors.forename: Wenhui
authors.initials: W
authors.lastname: Liu
authors.forename: Qi
authors.initials: Q
authors.lastname: Shen
authors.forename: Hongying
authors.initials: H
authors.lastname: Sun
authors.forename: Shuqing
authors.initials: S
authors.lastname: Chen
mesh_terms: D017667:Adipocytes / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D015870:Gene Expression; D019382:Human Growth Hormone / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D006801:Humans; D008066:Lipolysis / Q000187:drug effects*; D008297:Male; D010843:Pichia / Q000235:genetics*; D015202:Protein Engineering; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D051381:Rats; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology*; D012709:Serum Albumin / Q000235:genetics* / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; D012709:Serum Albumin; D019382:Human Growth Hormone
keywords: affinity chromatography; fusion protein; human growth hormone; lipolytic activity; protein purification