Photoprotection effect of the hyaluronic acid and ectoine composition was studied by evaluated its effects on inhibiting oxidative stress and MMP-1 production,ROS scavenging ability and promoting collagen synthesis in...Photoprotection effect of the hyaluronic acid and ectoine composition was studied by evaluated its effects on inhibiting oxidative stress and MMP-1 production,ROS scavenging ability and promoting collagen synthesis in skin cells under blue light irradiation.The result show that the composition is non-toxic to keratinocytes within a concentration of 5%.And the sample can significantly inhibit the increase of reactive oxygen species(ROS)caused by blue light irradiation at a concentration of 2%,and can also repaire the damaged skin by scavenge oxygen free radicals.At the same time,the composition can inhibit the increase of MMP-1 caused by blue light irradiation,so as to have the effect of inhibiting the decomposition of collagen and promoting the secretion of Collagen I.展开更多
Ectoine is a natural macromolecule protector and synthesized by some extremophiles.It provides protections against radiation-mediated oxidative damages and is widely used as a bioactive ingredient in pharmaceutics and...Ectoine is a natural macromolecule protector and synthesized by some extremophiles.It provides protections against radiation-mediated oxidative damages and is widely used as a bioactive ingredient in pharmaceutics and cosmetics.To meet its growing commercial demands,we engineered Escherichia coli strains for the high-yield production of ectoine.The ectABC gene cluster from the native ectoine producer Halomonas elongata was intro-duced into different Escherichia coli(E.Coil)strains via plasmids and 0.8 g L^(-1)of ectoine was produced in flask cultures by engineered E.coli BL21(DE3).Subsequently,we designed the ribosome-binding sites of the gene cluster to fine-tune the expressions of genes ectA,ectB,and ectC,which increased the ectoine yield to 1.6 g L^(-1).After further combinatorial overexpression of Corynebacterium glutamicum aspartate kinase mutant(G1A,C932T)and the H.elongate aspartate-semialdehyde dehydrogenase to increase the supply of the precursor,the titer of ectoine reached to 5.5 g L^(-1)in flask cultures.Finally,the engineered strain produced 60.7 g L^(-1)ectoine in fed-batch cultures with a conversion rate of 0.25 g/g glucose.展开更多
Ectoine is a high-value stabilizer and protective agent with various applications in enzyme industry,cosmetics,and biomedi-cine.In this study,rational engineering strategies have been implemented in Escherichia coli t...Ectoine is a high-value stabilizer and protective agent with various applications in enzyme industry,cosmetics,and biomedi-cine.In this study,rational engineering strategies have been implemented in Escherichia coli to efficiently produce ectoine.First,the synthetic pathway of ectoine was constructed in E.coli MG1655 by introducing an artificial thermal switch system harboring the ectABC cluster from Halomonas elongate,and the resulting strain produced 1.95 g/L ectoine.Second,crr encoding the glucose-specific enzyme II domain A of phosphotransferase system and iclR encoding the glyoxylate shunt transcriptional repressor were deleted in E.coli for enhancing the oxaloacetate supply,leading to the increasement of the ectoine titer to 9.09 g/L.Third,thrA encoding the bifunctional aspartokinase/homoserine dehydrogenase was removed from the genome to weaken the competitive pathway;simultaneously,an endogenous feedback-resistant lysC was overexpressed to complement the enzymatic activity deficiency of the aspartate kinase,leading to 30.36%increase of ectoine titer.Next,the expression of phosphoenolpyruvate carboxylase was modulated with varying gradient strength promoters to accelerate the biosynthesis efficiency of ectoine.Finally,aspDH encoding aspartate dehydrogenase from Pseudomonas aeruginosa PAO1 was overexpressed to further improve the biosynthesis of ectoine.The final strain MWZ003/pFT28-ectABC-EclysC^(*)-aspDH-ppc3 produced 30.37 g/L ectoine after 36-h fed-batch fermentation with a yield of 0.132 g/g glucose and a productivity of 0.844 g/(L h).展开更多
文摘Photoprotection effect of the hyaluronic acid and ectoine composition was studied by evaluated its effects on inhibiting oxidative stress and MMP-1 production,ROS scavenging ability and promoting collagen synthesis in skin cells under blue light irradiation.The result show that the composition is non-toxic to keratinocytes within a concentration of 5%.And the sample can significantly inhibit the increase of reactive oxygen species(ROS)caused by blue light irradiation at a concentration of 2%,and can also repaire the damaged skin by scavenge oxygen free radicals.At the same time,the composition can inhibit the increase of MMP-1 caused by blue light irradiation,so as to have the effect of inhibiting the decomposition of collagen and promoting the secretion of Collagen I.
基金supported by the National Key Research and Development Program of China(2021YFC2100800)the Jiangsu Province Natural Science Fund for Distinguished Young Scholars(BK20200025)+2 种基金the National Natural Science Foundation of China(32000058)a grant from the Key Technologies R&D Program of Jiangsu Province(BE2019630)the National First-class Discipline Program of Light Industry Technology and Engineering(LITE2018-16).
文摘Ectoine is a natural macromolecule protector and synthesized by some extremophiles.It provides protections against radiation-mediated oxidative damages and is widely used as a bioactive ingredient in pharmaceutics and cosmetics.To meet its growing commercial demands,we engineered Escherichia coli strains for the high-yield production of ectoine.The ectABC gene cluster from the native ectoine producer Halomonas elongata was intro-duced into different Escherichia coli(E.Coil)strains via plasmids and 0.8 g L^(-1)of ectoine was produced in flask cultures by engineered E.coli BL21(DE3).Subsequently,we designed the ribosome-binding sites of the gene cluster to fine-tune the expressions of genes ectA,ectB,and ectC,which increased the ectoine yield to 1.6 g L^(-1).After further combinatorial overexpression of Corynebacterium glutamicum aspartate kinase mutant(G1A,C932T)and the H.elongate aspartate-semialdehyde dehydrogenase to increase the supply of the precursor,the titer of ectoine reached to 5.5 g L^(-1)in flask cultures.Finally,the engineered strain produced 60.7 g L^(-1)ectoine in fed-batch cultures with a conversion rate of 0.25 g/g glucose.
基金This study was supported by the National Key R&D Program of China(2018YFA0900300)the National First-class Discipline Program of Light Industry Technology and Engineering(LITE2018-10)+1 种基金the Key Technology Project of Inner Mongolia Autonomous Region in China(2019GG302)the Collaborative Innovation Center of Jiangsu Modern Industrial Fermentation.
文摘Ectoine is a high-value stabilizer and protective agent with various applications in enzyme industry,cosmetics,and biomedi-cine.In this study,rational engineering strategies have been implemented in Escherichia coli to efficiently produce ectoine.First,the synthetic pathway of ectoine was constructed in E.coli MG1655 by introducing an artificial thermal switch system harboring the ectABC cluster from Halomonas elongate,and the resulting strain produced 1.95 g/L ectoine.Second,crr encoding the glucose-specific enzyme II domain A of phosphotransferase system and iclR encoding the glyoxylate shunt transcriptional repressor were deleted in E.coli for enhancing the oxaloacetate supply,leading to the increasement of the ectoine titer to 9.09 g/L.Third,thrA encoding the bifunctional aspartokinase/homoserine dehydrogenase was removed from the genome to weaken the competitive pathway;simultaneously,an endogenous feedback-resistant lysC was overexpressed to complement the enzymatic activity deficiency of the aspartate kinase,leading to 30.36%increase of ectoine titer.Next,the expression of phosphoenolpyruvate carboxylase was modulated with varying gradient strength promoters to accelerate the biosynthesis efficiency of ectoine.Finally,aspDH encoding aspartate dehydrogenase from Pseudomonas aeruginosa PAO1 was overexpressed to further improve the biosynthesis of ectoine.The final strain MWZ003/pFT28-ectABC-EclysC^(*)-aspDH-ppc3 produced 30.37 g/L ectoine after 36-h fed-batch fermentation with a yield of 0.132 g/g glucose and a productivity of 0.844 g/(L h).