Overexpression and characterization of two types of nitrile hydratases from Rhodococcus rhodochrous J1.
Nitrile hydratase (NHase) from Rhodococcus rhodochrous J1 is widely used for industrial production of acrylamide and nicotinamide. However, the two types of NHases (L-NHase and H-NHase) from R. rhodochrous J1 were only slightly expressed in E. coli by routine methods, which limits the comprehensive...
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| Format: | Article |
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Public Library of Science (PLoS)
2017-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC5482477?pdf=render |
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| author | Yao Lan Xiaohuan Zhang Zhongmei Liu Li Zhou Ruihua Shen Xianping Zhong Wenjing Cui Zhemin Zhou |
| author_facet | Yao Lan Xiaohuan Zhang Zhongmei Liu Li Zhou Ruihua Shen Xianping Zhong Wenjing Cui Zhemin Zhou |
| author_sort | Yao Lan |
| collection | DOAJ |
| description | Nitrile hydratase (NHase) from Rhodococcus rhodochrous J1 is widely used for industrial production of acrylamide and nicotinamide. However, the two types of NHases (L-NHase and H-NHase) from R. rhodochrous J1 were only slightly expressed in E. coli by routine methods, which limits the comprehensive and systematic characterization of the enzyme properties. We successfully expressed the two types of recombinant NHases in E. coli by codon-optimization, engineering of Ribosome Binding Site (RBS) and spacer sequences. The specific activity of the purified L-NHase and H-NHase were 400 U/mg and 234 U/mg, respectively. The molecular mass of L-NHase and H-NHase was identified to be 94 kDa and 504 kDa, respectively, indicating that the quaternary structure of the two types of NHases was the same as those in R. rhodochrous J1. H-NHase exhibited higher substrate and product tolerance than L-NHase. Moreover, higher activity and shorter culture time were achieved in recombinant E. coli, and the whole cell catalyst of recombinant E. coli harboring H-NHase has equivalent efficiency in tolerance to the high-concentration product relative to that in R. rhodochrous J1. These results indicate that biotransformation of nitrile by R. rhodochrous J1 represents a potential alternative to NHase-producing E. coli. |
| first_indexed | 2024-12-22T07:12:17Z |
| format | Article |
| id | doaj.art-d918ea969b19438a984236a19367f0de |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-22T07:12:17Z |
| publishDate | 2017-01-01 |
| publisher | Public Library of Science (PLoS) |
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| series | PLoS ONE |
| spelling | doaj.art-d918ea969b19438a984236a19367f0de2022-12-21T18:34:29ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01126e017983310.1371/journal.pone.0179833Overexpression and characterization of two types of nitrile hydratases from Rhodococcus rhodochrous J1.Yao LanXiaohuan ZhangZhongmei LiuLi ZhouRuihua ShenXianping ZhongWenjing CuiZhemin ZhouNitrile hydratase (NHase) from Rhodococcus rhodochrous J1 is widely used for industrial production of acrylamide and nicotinamide. However, the two types of NHases (L-NHase and H-NHase) from R. rhodochrous J1 were only slightly expressed in E. coli by routine methods, which limits the comprehensive and systematic characterization of the enzyme properties. We successfully expressed the two types of recombinant NHases in E. coli by codon-optimization, engineering of Ribosome Binding Site (RBS) and spacer sequences. The specific activity of the purified L-NHase and H-NHase were 400 U/mg and 234 U/mg, respectively. The molecular mass of L-NHase and H-NHase was identified to be 94 kDa and 504 kDa, respectively, indicating that the quaternary structure of the two types of NHases was the same as those in R. rhodochrous J1. H-NHase exhibited higher substrate and product tolerance than L-NHase. Moreover, higher activity and shorter culture time were achieved in recombinant E. coli, and the whole cell catalyst of recombinant E. coli harboring H-NHase has equivalent efficiency in tolerance to the high-concentration product relative to that in R. rhodochrous J1. These results indicate that biotransformation of nitrile by R. rhodochrous J1 represents a potential alternative to NHase-producing E. coli.http://europepmc.org/articles/PMC5482477?pdf=render |
| spellingShingle | Yao Lan Xiaohuan Zhang Zhongmei Liu Li Zhou Ruihua Shen Xianping Zhong Wenjing Cui Zhemin Zhou Overexpression and characterization of two types of nitrile hydratases from Rhodococcus rhodochrous J1. PLoS ONE |
| title | Overexpression and characterization of two types of nitrile hydratases from Rhodococcus rhodochrous J1. |
| title_full | Overexpression and characterization of two types of nitrile hydratases from Rhodococcus rhodochrous J1. |
| title_fullStr | Overexpression and characterization of two types of nitrile hydratases from Rhodococcus rhodochrous J1. |
| title_full_unstemmed | Overexpression and characterization of two types of nitrile hydratases from Rhodococcus rhodochrous J1. |
| title_short | Overexpression and characterization of two types of nitrile hydratases from Rhodococcus rhodochrous J1. |
| title_sort | overexpression and characterization of two types of nitrile hydratases from rhodococcus rhodochrous j1 |
| url | http://europepmc.org/articles/PMC5482477?pdf=render |
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