Using cDNA-AFLP technique, a specific fragment was isolated from cucumber cultivar Changchun mici possessing chilling tolerance induced at low temperature （15℃）. This fragment, named cctr 132, could not be induced ...Using cDNA-AFLP technique, a specific fragment was isolated from cucumber cultivar Changchun mici possessing chilling tolerance induced at low temperature （15℃）. This fragment, named cctr 132, could not be induced in the chilling sensitive cucumber cultivar Beijing jietou. After recovering the fragment, sequencing and translating, the results of blastx and blastp in GenBank of NCBI indicated that CCTR132 had 88.37% identities and 100% positives with Oryza sativa putative lysine decarboxylase-like protein respectively, and PGGXGTXXE, the putative conserved domain of lysine decarboxylase family, was detected from CCTR132, suggesting the cucumber chilling tolerance during germination is related to the expression of the lysine decarboxylase gene.展开更多
Cadaverine is the key monomer for the synthesis of nylon 5X.Efficient and alkaline stable lysine decarboxylases are highly desirable for cadaverine production as the reaction pH increasing from 6.3 to 8.5.However,the ...Cadaverine is the key monomer for the synthesis of nylon 5X.Efficient and alkaline stable lysine decarboxylases are highly desirable for cadaverine production as the reaction pH increasing from 6.3 to 8.5.However,the most studied lysine decarboxylase CadA(E.coli)lost almost all activity at pH 8.0,which is the foremost challenge for the industrial-cadaverine production.In this study,we first found that the Na^(+)-microenvironment significantly improved the alkaline stability of the disulfide engineered lysine decarboxylaseΔLdcEt3(P233C/L628C)(half-life 362 h),compared to the conventional buffer(half-life 0.66 h)at pH 8.0.Meanwhile,the whole-cell conversion efficiency of the industrial-grade L-lysine withΔLdcEt3 could reach up to 99%in 2 h in the fermenter.Experi-mental investigation and molecular dynamics confirmed that Na^(+)-microenvironment could improve active-aggregation state and affect secondary structure ofΔLdcEt3.Therefore,Na^(+)-microenvironment stabilizesΔLdcEt3 providing a great potential industrial application for high-level cadaverine production.展开更多
基金This work was partially supported by grants from National Hi-11ech R&D Program(2004241120)the National Natural Science Foundation of China(30470120)Ministry of Agriculture Vegetable Genetic and Physiological Key Laboratory.
文摘Using cDNA-AFLP technique, a specific fragment was isolated from cucumber cultivar Changchun mici possessing chilling tolerance induced at low temperature （15℃）. This fragment, named cctr 132, could not be induced in the chilling sensitive cucumber cultivar Beijing jietou. After recovering the fragment, sequencing and translating, the results of blastx and blastp in GenBank of NCBI indicated that CCTR132 had 88.37% identities and 100% positives with Oryza sativa putative lysine decarboxylase-like protein respectively, and PGGXGTXXE, the putative conserved domain of lysine decarboxylase family, was detected from CCTR132, suggesting the cucumber chilling tolerance during germination is related to the expression of the lysine decarboxylase gene.
基金supported by the National Natural Science Foundation of China (grant number 22078346)Beijing Nova Program of Science and Technology (Z201100006820141)+4 种基金Innovation Academy for Green Manufacture, CAS (IAGM2020C19)Natural Science Foundation of Beijing (2204097)Henan Key Research and Development Project (202102210046)Hebei Provincial Natural Science Foundation (B2020103010)the CAS Pioneer Hundred Program。
文摘Cadaverine is the key monomer for the synthesis of nylon 5X.Efficient and alkaline stable lysine decarboxylases are highly desirable for cadaverine production as the reaction pH increasing from 6.3 to 8.5.However,the most studied lysine decarboxylase CadA(E.coli)lost almost all activity at pH 8.0,which is the foremost challenge for the industrial-cadaverine production.In this study,we first found that the Na^(+)-microenvironment significantly improved the alkaline stability of the disulfide engineered lysine decarboxylaseΔLdcEt3(P233C/L628C)(half-life 362 h),compared to the conventional buffer(half-life 0.66 h)at pH 8.0.Meanwhile,the whole-cell conversion efficiency of the industrial-grade L-lysine withΔLdcEt3 could reach up to 99%in 2 h in the fermenter.Experi-mental investigation and molecular dynamics confirmed that Na^(+)-microenvironment could improve active-aggregation state and affect secondary structure ofΔLdcEt3.Therefore,Na^(+)-microenvironment stabilizesΔLdcEt3 providing a great potential industrial application for high-level cadaverine production.
