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Expression, purification, and lipolytic activity of recombinant human serum albumin fusion proteins with one domain of human growth hormone in P ichia pastoris
Author(s) -
Wang Furong,
Wu Min,
Liu Wenhui,
Shen Qi,
Sun Hongying,
Chen Shuqing
Publication year - 2013
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.1108
Subject(s) - pichia pastoris , fusion protein , human serum albumin , recombinant dna , biochemistry , western blot , fusion gene , albumin , biology , serum albumin , chemistry , microbiology and biotechnology , gene
Abstract Human growth hormone (h GH ) 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 h GH . The amino terminus of h GH 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 h GH were constructed and expressed in Pichia pastoris . The recombinant proteins were purified and characterized by SDS ‐ PAGE and W estern blot. The preliminary stability tests demonstrated that HSA –h GH 166–191 and HSA –h GH 177–191 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.