文献类型：期刊文献

英文题名：Efficient production of α-L-rhamnosidase podoRha through multi-strategy synergistic regulation involving molecular chaperones, osmolytes, and high-density fermentation management in Escherichia coli, and its application in isoquercitrin production via resting cell transformation

作者：Xie, Jingcong[1,2,3] Zhao, Jian[1,2,3] Zhang, Ning[1,2,3] Xu, Hao[1,2,3] Yang, Jing[1,2,3] Tao, Ran[1,2,3] Jiang, Jianchun[1,2,3]

第一作者：Xie, Jingcong

通信作者：Zhang, N[1];Jiang, JC[1];Zhang, N[2];Jiang, JC[2];Zhang, N[3];Jiang, JC[3]

机构：[1]Chinese Acad Forestry, Inst Chem Ind Forest Prod, Natl Key Lab Dev & Utilizat Forest Food Resources, Nanjing 210042, Peoples R China;[2]Natl Forestry & Grassland Adm, Natl Engn Res Ctr Low Carbon Proc & Utilizat Fores, Key Lab Biomass Energy & Mat, Key Lab Chem Engn Forest Prod, Nanjing 210042, Jiangsu, Peoples R China;[3]Nanjing Forestry Univ, Int Innovat Ctr Forest Chem & Mat, Jiangsu Coinnovat Ctr Efficient Proc & Utilizat Fo, Nanjing 210037, Peoples R China

年份：2025

卷号：320

外文期刊名：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES

收录：;EI(收录号：20252718728703);Scopus(收录号：2-s2.0-105009697703);WOS:【SCI-EXPANDED(收录号:WOS:001531394200023)】；

基金：This work was financially supported by Fundamental Research funds for the Central Nonprofit Research Institution of CAF (Grant No. CAFYBB2021ZA003) , National Natural Science Foundation of China (No. 32101477) , Jiangsu Key Laboratory of Biomass Energy and Mate-rial (No. JSBEM-S-201802) , National Natural Science Foundation of China (No. 32401530) .

语种：英文

外文关键词：Isoquercitrin transformation; Molecular chaperone and osmolytes; High-density fermentation of E. coli

摘要：The alpha-L-rhamnosidase podoRha could selectively cleaves the alpha-1,6 glycosidic bond, exhibiting remarkable activity in the bioconversion of rutin to isoquercitrin. In order to establish a straightforward and efficient isoquercitrin production system, a multi-strategy combination production approach was employed. This approach involved the co-expression of a molecular chaperone (DnaK-DnaJ-DrpE), co-cultivation with osmolytes (betaine), and high-density fermentation. An alpha-L- rhamnosidase activity of 407.15 U center dot mL- 1 was achieved in a 7-L bioreactor, representing the highest alpha-L-rhamnosidase activity reported to date. Furthermore, a recombinant podoRha productivity of 10.323 kU center dot g-1 was observed, which was 1.50-fold higher than that of BL21-podoRha in a 7-L bioreactor without osmolytes. Therefore, this approach succeeds to significantly enhance both recombinant podoRha productivity and enzyme activity by synergistically reducing the formation of inclusion bodies and improving soluble recombinant podoRha expression. Importantly, it also was found that the batch addition of rutin in a resting cell transformation system facilitated the enhancement of transformation efficiency. This approach led to the efficient conversion of 8 g center dot L- 1 of rutin into 5.89 g center dot L- 1 of isoquercitrin, achieving a molar transformation rate of 96.88 %. This production scheme is anticipated to offer an efficient and cost-effective strategy for the overproduction of recombinant podoRha, enabling the efficient production of highly active natural products.