Novel system to produce highly functional proteins as protein inclusion in Escherichia coli.

2016 Fiscal Year Final Research Report

• Project/Area Number: 26660268
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Insect science
• Research Institution: Okayama University
• Principal Investigator: Hayakawa Toru 岡山大学, 自然科学研究科, 助教 (30313555)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: Bacillus thuringiensis / 4AaCter-tag / Cry4Aa / Cry46Ab / 大腸菌 / タンパク質凝集体形成

Outline of Final Research Achievements

4AaCter is a peptide-tag derived from mosquitocidal Cry4Aa toxin, and fusion with this tag facilitate formation of the protein inclusion in Escherichia coli. In this study, through the analyses using various truncated mutants of 4AaCter-tag, two domains (MIF1 and 2) that related to formation of protein inclusion were identified in the amino acid stretch of 4AaCter. Our data suggested that MIF domains polymerize by binding itself and/or each other, and facilitate formation of protein inclusion. In addition, proteins (designated as P49 and P50) which may be related to the formation of protein inclusion were detected in Escherichia coli proteins. In this study, the potency of 4AaCter-tag was investigated to prepare the formulation of insecticidal Cry46Ab toxin. Application of 4AaCter-tag yielded excellent results, including increase in the production level, simplification of the product purification and high toxicity against Culex pipiens larvae.

Free Research Field

境界農学・昆虫科学