Numerous studies have shown that intestinal and urinary tract flora are closely related to the formation of kidney stones.The removal of probiotics represented by lactic acid bacteria and the colonization of pathogeni...Numerous studies have shown that intestinal and urinary tract flora are closely related to the formation of kidney stones.The removal of probiotics represented by lactic acid bacteria and the colonization of pathogenic bacteria can directly or indirectly promote the occurrence of kidney stones.However,currently existing natural probiotics have limitations.Synthetic biology is an emerging discipline in which cells or living organisms are genetically designed and modified to have biological functions that meet human needs,or even create new biological systems,and has now become a research hotspot in various fields.Using synthetic biology approaches of microbial engineering and biological redesign to enable probiotic bacteria to acquire new phenotypes or heterologous protein expression capabilities is an important part of synthetic biology research.Synthetic biology modification of microorganisms in the gut and urinary tract can effectively inhibit the development of kidney stones by a range of means,including direct degradation of metabolites that promote stone production or indirect regulation of flora homeostasis.This article reviews the research status of engineered microorganisms in the prevention and treatment of kidney stones,to provide a new and effective idea for the prevention and treatment of kidney stones.展开更多
A azoreductase gene with 537 bp was obtained by PCR amplification from Rhodobacter sphaeroides AS1 1737 The enzyme, with a molecular weight of 18 7 kD, was efficiently expressed in Escherichia coli and its biodegr...A azoreductase gene with 537 bp was obtained by PCR amplification from Rhodobacter sphaeroides AS1 1737 The enzyme, with a molecular weight of 18 7 kD, was efficiently expressed in Escherichia coli and its biodegradation characteristics for azo dyes were investigated. Furthermore, the reaction kinetics and mechanism of azo dyes catalyzed by the genetically engineered azoreductase were studied in detail. The presence of a hydrazo-intermediate was identified, which provided a convincing evidence for the assumption that azo dyes were degraded via an incomplete reduction stage.展开更多
Engineering microorganisms to withstand extreme temperatures(>80℃)remains a critical challenge in industrial biotechnology owing to limited genetic tools and poor mechanistic understanding of microbial thermoadapt...Engineering microorganisms to withstand extreme temperatures(>80℃)remains a critical challenge in industrial biotechnology owing to limited genetic tools and poor mechanistic understanding of microbial thermoadapta-tion.We aimed to develop a novel Geobacillus stearothermophilus strain with remarkable thermal resilience through an integrated approach combining adaptive laboratory evolution and rational genetic engineering.Progressive thermal adaptation(70-80℃)followed by genome reduction generated a mutant(SL-1-80)with enhanced sta-bility at 80℃.Subsequent combinatorial overexpression of eight heat-associated genes(murD,cysM,grpE,groES,hsp33,hslO,hrcA,clpE)synergistically extended its survival to 85℃.Genomic and transcriptomic analyses re-vealed a triple mechanism:(1)strategic deletion of transposable elements(IS5377/IS4/IS110)reduced genomic instability,(2)co-activation of chaperone systems(GroES-GrpE)and redox homeostasis enzymes(HslO-Hsp33)enhanced protein folding and oxidative stress resistance,and(3)metabolic plasticity(BglG and HTH-domain transcriptional repressor),motility optimization(FliY),and transcriptional reprogramming(Sigma-D,DUF47-family chaperone and HTH-domain transcriptional repressor)facilitated nutrient acquisition and motility-based environmental navigation under stress.Furthermore,we established the first high-efficiency electroporation pro-tocol(10^(4) transformants/μg DNA)for this genus,enabling ATP-enhanced heterologous protein expression under heat stress.This study provided a robust platform organism for high-temperature bioprocessing and a mecha-nistic blueprint for engineering microbial thermotolerance,addressing key limitations in applications such as microbial-enhanced oil recovery and industrial enzyme production.展开更多
文摘Numerous studies have shown that intestinal and urinary tract flora are closely related to the formation of kidney stones.The removal of probiotics represented by lactic acid bacteria and the colonization of pathogenic bacteria can directly or indirectly promote the occurrence of kidney stones.However,currently existing natural probiotics have limitations.Synthetic biology is an emerging discipline in which cells or living organisms are genetically designed and modified to have biological functions that meet human needs,or even create new biological systems,and has now become a research hotspot in various fields.Using synthetic biology approaches of microbial engineering and biological redesign to enable probiotic bacteria to acquire new phenotypes or heterologous protein expression capabilities is an important part of synthetic biology research.Synthetic biology modification of microorganisms in the gut and urinary tract can effectively inhibit the development of kidney stones by a range of means,including direct degradation of metabolites that promote stone production or indirect regulation of flora homeostasis.This article reviews the research status of engineered microorganisms in the prevention and treatment of kidney stones,to provide a new and effective idea for the prevention and treatment of kidney stones.
文摘A azoreductase gene with 537 bp was obtained by PCR amplification from Rhodobacter sphaeroides AS1 1737 The enzyme, with a molecular weight of 18 7 kD, was efficiently expressed in Escherichia coli and its biodegradation characteristics for azo dyes were investigated. Furthermore, the reaction kinetics and mechanism of azo dyes catalyzed by the genetically engineered azoreductase were studied in detail. The presence of a hydrazo-intermediate was identified, which provided a convincing evidence for the assumption that azo dyes were degraded via an incomplete reduction stage.
基金supported by the National Key R&D Program of China(grant number 2023YFC3402003)the National Natural Science Foundation of Shandong province(grant number ZR2023YQ028)+4 种基金Tais-han Scholar Program of Shandong Province in China(grant number tsqn202312007)Guangdong Basic and Applied Basic Research Foun-dation(grant number 2022A1515110795)the Recruitment Program of Global Experts(grant number 1000 Plan)the Program of Introduc-ing Talents of Discipline to Universities(grant number B16030)SKLMT Frontiers and Challenges Project of Shandong University.
文摘Engineering microorganisms to withstand extreme temperatures(>80℃)remains a critical challenge in industrial biotechnology owing to limited genetic tools and poor mechanistic understanding of microbial thermoadapta-tion.We aimed to develop a novel Geobacillus stearothermophilus strain with remarkable thermal resilience through an integrated approach combining adaptive laboratory evolution and rational genetic engineering.Progressive thermal adaptation(70-80℃)followed by genome reduction generated a mutant(SL-1-80)with enhanced sta-bility at 80℃.Subsequent combinatorial overexpression of eight heat-associated genes(murD,cysM,grpE,groES,hsp33,hslO,hrcA,clpE)synergistically extended its survival to 85℃.Genomic and transcriptomic analyses re-vealed a triple mechanism:(1)strategic deletion of transposable elements(IS5377/IS4/IS110)reduced genomic instability,(2)co-activation of chaperone systems(GroES-GrpE)and redox homeostasis enzymes(HslO-Hsp33)enhanced protein folding and oxidative stress resistance,and(3)metabolic plasticity(BglG and HTH-domain transcriptional repressor),motility optimization(FliY),and transcriptional reprogramming(Sigma-D,DUF47-family chaperone and HTH-domain transcriptional repressor)facilitated nutrient acquisition and motility-based environmental navigation under stress.Furthermore,we established the first high-efficiency electroporation pro-tocol(10^(4) transformants/μg DNA)for this genus,enabling ATP-enhanced heterologous protein expression under heat stress.This study provided a robust platform organism for high-temperature bioprocessing and a mecha-nistic blueprint for engineering microbial thermotolerance,addressing key limitations in applications such as microbial-enhanced oil recovery and industrial enzyme production.
