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Precise l-threonine-to-l-isoleucine pathway regulation for engineering high-efficiency whole-cell biocatalysts 认领 引用 被引量：1
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作者 Heng Zhang Fuqiang Song +4 位作者 Ke Wang Faqing Wu Lihao Deng Kun Qiu Jingwen Zhou 《Synthetic and Systems Biotechnology》 SCIE CSCD 2026年第2期1-9,共9页
l-Isoleucine(L-Ile),a critical branched-chain amino acid with diverse applications in food,pharmaceutical,and cosmetic industries,is difficult to produce efficiently at scale in microbial systems due to metabolic bott... l-Isoleucine(L-Ile),a critical branched-chain amino acid with diverse applications in food,pharmaceutical,and cosmetic industries,is difficult to produce efficiently at scale in microbial systems due to metabolic bottlenecks and cofactor limitations.This study metabolically engineered Escherichia coli BL21(DE3)to develop a whole-cell biocatalyst for efficient L-Ile biosynthesis.Key strategies included screening acetohydroxy acid synthase(AHAS)isoenzymes,identifying ilvGM-encoded AHAS II as the optimal enzyme,relieving feedback inhibition of ilvA(encoding l-threonine dehydratase)through mutant screening,and optimizing genetic circuits(promoter tuning,plasmid copy number).Dual-precursor supplementation revealed l-threonine as a critical factor for suppressing l-valine byproduct.Fed-batch fermentation in a 5 L bioreactor achieved a peak molar conversion rate of 98.4%,yielding 40.1 g/L L-Ile within 36 h.The mass conversion rate(L-Ile/glucose)achieved 0.36 g/g and the production efficiency achieved 1.11 g/L/h,demonstrating the feasibility of whole-cell catalysis.This work provides a robust framework for industrial L-Ile production and transferable strategies for branched-chain amino acid pathway optimization. 展开更多
关键词 l-isoleucine production Whole-cell biocatalyst Transcription regulation Metabolic engineering Acetohydroxy acid synthase Dual-precursor supplementation
Cloning and analysis of a cDNA encoding acetohydroxy acid isomeroreductase from G2 pea 认领 引用 被引量：3
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作者 XU Huasong XU Yunjian +3 位作者 GU Xuesong HU Tingzhang SU Yanhui ZHU Yuxian 《Chinese Science Bulletin》 2001年第21期1808-1812,共5页
Using cDNA representational difference analysis (cDNA RDA) method, we have successfully isolated a gene fragment whose expression was specifically induced by external GAa application. Screening a G2 pea cDNA library u... Using cDNA representational difference analysis (cDNA RDA) method, we have successfully isolated a gene fragment whose expression was specifically induced by external GAa application. Screening a G2 pea cDNA library using this fragment as a probe, we obtained a 2036 bp full-length cDNA. It contains a 1746 bp open reading frame and encodes a protein of 581 amino acids with a theoretical molecular weight of 64 ku. It shares high-level sequenceidentity with AAIR genes from other plant species. This cDNA was cloned into expression vector and recombinant E. coli DH5α cells with remarkable AAIR enzyme activity were obtained. 展开更多
关键词 G2 pea clone acetohydroxy acid isomeroreductase prokaryotic expression.
Syntheses,Crystal Structures and Bioactivities of Two Novel Isatin Derivatives 认领 引用
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作者 SHANG Jian-li LI Hui-dong +3 位作者 SHANG Jun SONG Hai-bjn LI Zheng-ming WANG Jian-guo 《Chemical Research in Chinese Universities》 SCIE CAS CSCD 2011年第4期595-598,共4页
Two novel compounds 1-(4-fluorobenzyl)-4-chloro-(Z)-3-benzoylhydrazono-2-indolinone(1)and 1-(4-methoxybenzyl)-(Z)-3-benzoylhydrazono-2-indolinone(2)were synthesized and their crystal structures were determined by sing... Two novel compounds 1-(4-fluorobenzyl)-4-chloro-(Z)-3-benzoylhydrazono-2-indolinone(1)and 1-(4-methoxybenzyl)-(Z)-3-benzoylhydrazono-2-indolinone(2)were synthesized and their crystal structures were determined by single-crystal X-ray diffraction.Compound 1(C22H15ClFN3O2)crystallized in the triclinic system,space group P1·with a=0.94198(19)nm,b=1.4339(3)nm,c=1.5018(3)nm,α=101.58(3)°,β=102.96(3)°,γ=102.73°,V=1.8602(6)nm3,Mr=407.82,Dc=1.456 g/cm3,μ=0.240 mm-1,F(000)=840,Z=4,R1=0.0442 and wR2=0.1064.Compound 2(C23H19N3O3)crystallized in the triclinic system,space group P1·with a=1.0022(2)nm,b=1.0192(2)nm,c=1.0461(2)nm,α=99.86(3)°,β=117.30(3)°,γ=94.13(3)°,V=0.9215(3)nm3,Mr=385.41,Dc=1.389 g/cm3,μ=0.094 mm-1,F(000)=404,Z=2,R1=0.0403 and wR2=0.1142.The preliminary herbicidal activities of the two compounds were also evaluated. 展开更多
关键词 Acetohydroxy acid synthase(AHAS) Crystal structure Isatin derivative Herbicidal activity
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