Impact of orientation and flexibility of peptide linkers on T-maritima lipase Tm1350 displayed on Bacillus subtilis spores surface using CotB as fusion partner | |
Ullah, Jawad1; Chen, Huayou1,2; Vastermark, Ake3,4; Jia, Jinru1; Wu, Bangguo1; Ni, Zhong1; Le, Yilin1; Wang, Hongcheng1 | |
2017-09-01 | |
Source Publication | WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
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ISSN | 0959-3993 |
Volume | 33Issue:9Pages:166 |
Abstract | Fusion protein construction often requires peptide linkers for prolonged conformation, extended stability and enzyme activity. In this study a series of fusion between Thermotoga maritima lipase Tm1350 and Bacillus subtillis coat protein CotB, comprising of several peptide linkers, with different length, flexibility and orientations were constructed. Effects of temperature, pH and chemicals were examined, on the activity of displayed enzyme. The fusion protein with longer flexible linkers L9 [(GGGGS)(4)] and L7 (GGGGS-GGGGS-EAAAK-EAAAK-GGGGS-GGGGS) possess 1.29 and 1.16-fold higher activity than the original, under optimum temperature and pH respectively. Moreover, spore surface displaying Tm1350 with L3 (EAAAK-GGGGS) and L9 ((GGGGS) 4) showed extended thermostably, maintaining 1.40 and 1.35-fold higher activity than the original respectively, at 80 degrees C after 5 h of incubation. The enzyme activity of linkers with different orientation, including L5, L6 and L7 was determined, where L7 maintained 1.05 and 1.27-fold higher activity than L5 and L6. Effect of 0.1% proteinase K, bromelain, 20% ethanol and 30% methanol was investigated. Linkers with appropriate Glycine residues (flexible) showed higher activity than Alanine residues (rigid). The activity of the displayed enzyme can be improved by maintaining orientation and flexibility of peptide linkers, to evaluate high activity and stability in industrial processes. |
Keyword | Bacillus Subtilis Enzyme Activity Linkers Surface Display Tm1350 |
Subtype | Article |
WOS Headings | Science & Technology ; Life Sciences & Biomedicine |
DOI | 10.1007/s11274-017-2327-1 |
Indexed By | SCI |
Language | 英语 |
WOS Keyword | DOMAIN LINKERS ; ANCHOR PROTEIN ; MSB8 NITRILASE ; COAT ; ACTIVATION ; CARRIER ; ENZYME ; DESIGN ; SYSTEM |
WOS Research Area | Biotechnology & Applied Microbiology |
WOS Subject | Biotechnology & Applied Microbiology |
Funding Organization | Open Funding Project of National Key Laboratory of Biochemical Engineering, The Key R&D Program of Jiangsu Province (Modern Agriculture), China(BE2017355) ; Open Funding Project of the State Key Laboratory of Bioreactor Engineering ; Agricultural Sci-Tech Self-Innovation Program of Jiangsu Province, China(CX(17)3044) ; JSPS fellowship(PE 16042) |
WOS ID | WOS:000411062900005 |
Citation statistics | |
Document Type | 期刊论文 |
Identifier | http://ir.ipe.ac.cn/handle/122111/23228 |
Collection | 生化工程国家重点实验室 |
Affiliation | 1.Jiangsu Univ, Inst Life Sci, Zhenjiang 212000, Jiangsu, Peoples R China 2.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China 3.Univ Calif San Diego, Div Biol Sci, La Jolla, CA 92093 USA 4.Nitech, Showa Ku, Nagoya, Aichi 4668555, Japan |
Recommended Citation GB/T 7714 | Ullah, Jawad,Chen, Huayou,Vastermark, Ake,et al. Impact of orientation and flexibility of peptide linkers on T-maritima lipase Tm1350 displayed on Bacillus subtilis spores surface using CotB as fusion partner[J]. WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY,2017,33(9):166. |
APA | Ullah, Jawad.,Chen, Huayou.,Vastermark, Ake.,Jia, Jinru.,Wu, Bangguo.,...&Wang, Hongcheng.(2017).Impact of orientation and flexibility of peptide linkers on T-maritima lipase Tm1350 displayed on Bacillus subtilis spores surface using CotB as fusion partner.WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY,33(9),166. |
MLA | Ullah, Jawad,et al."Impact of orientation and flexibility of peptide linkers on T-maritima lipase Tm1350 displayed on Bacillus subtilis spores surface using CotB as fusion partner".WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY 33.9(2017):166. |
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Impact of orientatio(1665KB) | 期刊论文 | 出版稿 | 限制开放 | CC BY-NC-SA | Application Full Text |
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