Tags

Type your tag names separated by a space and hit enter

Enzymatic production of trans-4-hydroxy-l-proline by proline 4-hydroxylase.
Microb Biotechnol. 2020 Jul 03 [Online ahead of print]MB

Abstract

Trans-4-hydroxy-l-proline (Hyp) is a useful chiral building block for production of many nutritional supplements and pharmaceuticals. However, it is still challenging for industrial production of Hyp due to heavy environmental pollution and low production efficiency. To establish a green and efficient process for Hyp production, the proline 4-hydroxylase (DsP4H) from Dactylosporangium sp. RH1 was overexpressed and functionally characterized in Escherichia coli BL21(DE3). The recombinant DsP4H with l-proline as a substrate exhibited Km , kcat and kcat /Km values up to 0.80 mM, 0.52 s-1 and 0.65 s-1 ·mM-1 respectively. Furthermore, DsP4H showed the highest activity at 35°C and pH 6.5 towards l-proline. The highest enzyme activity of 175.6 U mg-1 was achieved by optimizing culture parameters. Under the optimal transformation conditions in a 5-l fermenter, Hyp titre, conversion rate and productivity were up to 99.9 g l-1 , 99.9% and 2.77 g l-1 h-1 respectively. This strategy described here provides an efficient method for production of Hyp and thus has a great potential in industrial application.

Authors+Show Affiliations

State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China. International Joint Laboratory on Food Safety, Jiangnan University, Wuxi, 214122, China.State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China. Shaoxing Baiyin Biotechnology Co. Ltd, Shaoxing, 312000, China.State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China. International Joint Laboratory on Food Safety, Jiangnan University, Wuxi, 214122, China.State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China.State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China. International Joint Laboratory on Food Safety, Jiangnan University, Wuxi, 214122, China.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32618422

Citation

Chen, Xiulai, et al. "Enzymatic Production of Trans-4-hydroxy-l-proline By Proline 4-hydroxylase." Microbial Biotechnology, 2020.
Chen X, Yi J, Liu J, et al. Enzymatic production of trans-4-hydroxy-l-proline by proline 4-hydroxylase. Microb Biotechnol. 2020.
Chen, X., Yi, J., Liu, J., Luo, Q., & Liu, L. (2020). Enzymatic production of trans-4-hydroxy-l-proline by proline 4-hydroxylase. Microbial Biotechnology. https://doi.org/10.1111/1751-7915.13616
Chen X, et al. Enzymatic Production of Trans-4-hydroxy-l-proline By Proline 4-hydroxylase. Microb Biotechnol. 2020 Jul 3; PubMed PMID: 32618422.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Enzymatic production of trans-4-hydroxy-l-proline by proline 4-hydroxylase. AU - Chen,Xiulai, AU - Yi,Juyang, AU - Liu,Jia, AU - Luo,Qiuling, AU - Liu,Liming, Y1 - 2020/07/03/ PY - 2020/04/03/received PY - 2020/05/23/revised PY - 2020/06/01/accepted PY - 2020/7/4/entrez JF - Microbial biotechnology JO - Microb Biotechnol N2 - Trans-4-hydroxy-l-proline (Hyp) is a useful chiral building block for production of many nutritional supplements and pharmaceuticals. However, it is still challenging for industrial production of Hyp due to heavy environmental pollution and low production efficiency. To establish a green and efficient process for Hyp production, the proline 4-hydroxylase (DsP4H) from Dactylosporangium sp. RH1 was overexpressed and functionally characterized in Escherichia coli BL21(DE3). The recombinant DsP4H with l-proline as a substrate exhibited Km , kcat and kcat /Km values up to 0.80 mM, 0.52 s-1 and 0.65 s-1 ·mM-1 respectively. Furthermore, DsP4H showed the highest activity at 35°C and pH 6.5 towards l-proline. The highest enzyme activity of 175.6 U mg-1 was achieved by optimizing culture parameters. Under the optimal transformation conditions in a 5-l fermenter, Hyp titre, conversion rate and productivity were up to 99.9 g l-1 , 99.9% and 2.77 g l-1 h-1 respectively. This strategy described here provides an efficient method for production of Hyp and thus has a great potential in industrial application. SN - 1751-7915 UR - https://www.unboundmedicine.com/medline/citation/32618422/Enzymatic_production_of_trans-4-hydroxy-l-proline_by_proline_4-hydroxylase L2 - https://doi.org/10.1111/1751-7915.13616 DB - PRIME DP - Unbound Medicine ER -
Try the Free App:
Prime PubMed app for iOS iPhone iPad
Prime PubMed app for Android
Prime PubMed is provided
free to individuals by:
Unbound Medicine.