Algae are reported to be corrosive,while little is known about the role of the algae associated bacteria in the corrosion process.In the present study,Halomonas titanicae was isolated from a culture of an alga strain,...Algae are reported to be corrosive,while little is known about the role of the algae associated bacteria in the corrosion process.In the present study,Halomonas titanicae was isolated from a culture of an alga strain,Spirulina platensis,and identified through 16 S rRNA gene analysis.Corrosion behavior of 304L stainless steel(SS)coupons in the presence and absence of H.titanicae was characterized by using electrochemical measurements and surface analysis.The results showed that H.titanicae significantly accelerated the corrosion rate and decreased the pitting potential of 304L SS in the biotic medium.After removal of the corrosion products and biofilms,severe pitting corrosion caused by H.titanicae was observed.The largest pit depth after 14 d reached 6.6μm,which was 5.5 times higher than that of the sterile control(1.2μm).This is the first report revealing that an alga associated bacterium can induce microbiologically influenced corrosion(MIC),and a further concern is raised that whether algae play a role in the MIC process.展开更多
In recent years,red tides occurred frequently in coastal areas worldwide.Various methods based on the use of clay,copper sulfate,and bacteria have been successful in controlling red tides to some extent.As a new defen...In recent years,red tides occurred frequently in coastal areas worldwide.Various methods based on the use of clay,copper sulfate,and bacteria have been successful in controlling red tides to some extent.As a new defensive agent,marine microorganisms are important sources of compounds with potent inhibitory bioactivities against red-tide microalgae,such as Gymnodinium sp.(Pyrrophyta).In this study,we isolated a marine bacterium,HSB07,from seawater collected from Hongsha Bay,Sanya,South China Sea.Based on its 16S rRNA gene sequence and biochemical characteristics,the isolated strain HSB07 was identified as a member of the genus Halomonas.A crude ethyl acetate extract of strain HSB07 showed moderate inhibition activity against Gymnodinium sp.in a bioactive prescreening experiment.The extract was further separated into fractions A,B,and C by silica gel column chromatography.Fractions B and C showed strong inhibition activities against Gymnodinium.This is the first report of inhibitory activity of secondary metabolites of a Halomonas bacterium against a red-tide-causing microalga.展开更多
Nitrification is a key step in the global nitrogen cycle.Compared with autotrophic nitrification,heterotrophic nitrification remains poorly understood.In this study,Halomonas venusta MA-ZP17-13,isolated from seawater ...Nitrification is a key step in the global nitrogen cycle.Compared with autotrophic nitrification,heterotrophic nitrification remains poorly understood.In this study,Halomonas venusta MA-ZP17-13,isolated from seawater in shrimp aquaculture (Penaeus vannamei),could simultaneously undertake nitrification and denitrification.With the initial ammonium concentration at 100 mg/L,the maximum ammonium-nitrogen removal rate reached98.7%under the optimal conditions including C/N concentration ratio at 5.95,p H at 8.93,and Na Cl at 2.33%.The corresponding average removal rate was 1.37 mg/(L·h)(according to nitrogen) in 3 d at 11.2℃.By whole genome sequencing and analysis,nitrification-and denitrification-related genes were identified,including ammonia monooxygenase,nitrate reductase,nitrite reductase,nitric oxide dioxygenase and nitric oxide synthase;while no gene encoding hydroxylamine oxidase was identified,it implied the existence of a novel nitrification pathway from hydroxylamine to nitrate.These results indicate heterotrophic bacterium H.venusta MA-ZP17-13 can undertake simultaneous nitrification and denitrification at low temperature and has potential for NH_(4)^(+)-N/NH_(3)-N removal in marine aquaculture systems.展开更多
Halomonas sp.YSR-3 was isolated from the Yellow Sea and identified as a polyphosphate-accumulating bacterium and the characteristics of its intracellular polyphosphate(polyP)granules and phosphorus absorption were stu...Halomonas sp.YSR-3 was isolated from the Yellow Sea and identified as a polyphosphate-accumulating bacterium and the characteristics of its intracellular polyphosphate(polyP)granules and phosphorus absorption were studied.Most YSR-3 cells stored one or two polyP granules in regular appearance and high-density.The diameter of the granules was about 400 nm measuring by a transmission electron microscope(TEM).After stained with 4,6-diamidino-2-phenylindole(DAPI)and visualized by a fluorescence microscope,the cells turned blue and the granules were bright yellow.The composition of granules includes P(major ingredient),Mg,S,K,and Ca as detected by an energy dispersive X-ray spectrometer(EDS).When inorganic phosphorus(po34-)and ferric ion(Fe3+)were added into media,the biomass increased and the cells formed intracellular polyP granules owing to the phosphorus assimilation from media.The YSR-3 obtained higher biomass by adding 0.02 g/L FePO4 than 0.005 g/L and 0.01 g/L FePO4;however,the phosphorus absorption was higher with 0.01 g/L FePO4 than 0.005 g/L and 0.02 g/L FePO4.The optical density at wavelength 480 nm(OD480nm)was 0.79 and 100%cells could form intracellular polyP granules.These results show that strain YSR-3 is able to acquire higher biomass and absorb more inorganic phosphorus when 0.01 g/L FePO4 is added.The characteristics of absorbing and storing phosphorus as intracellular inorganic polyP granules have a potential for application in high-efficiency phosphorus removal in wastewater treatment.展开更多
Hydantoinase is involved in the production of optically pure amino acids from racemic 5-mono-substituted hydantoins. We measured the D-hydantoinase activity in marine Halomonas sp. YSR-3 and amplified the D-hydantoina...Hydantoinase is involved in the production of optically pure amino acids from racemic 5-mono-substituted hydantoins. We measured the D-hydantoinase activity in marine Halomonas sp. YSR-3 and amplified the D-hydantoinase gene by PCR. The gene was inserted into vector pGM-T and transformed into E. coli TOP10. The positive transformants with the D-hydantoinase gene were sequenced. The sequenced fragment comprises 1 510 base pairs. The D-hydantoinase gene from YSR-3 is 77% similar to that from Pseudomonas entornophila L4 by searching against the NCBI databse. The protein product of the YSR-3 D-hydantoinase gene is 75%, 73%, and 70% similar to those from Pseudomonasfluorescens Pf-5, Marinornonas sp. MED121, and Burkholderia vietnamiensis G4, respectively. The difference of the D-hydantoinase gene between marine Halornonas sp. YSR-3 and other terrestrial organisms is distinct.展开更多
A short-rod-shaped halotolerant bacterium,designated CUG00002T,was isolated from the sediment of Xiaochaidan Lake of Qadam Basin,China,using R2A medium.The cells are Gram-negative,aerobic and forming creamy and circul...A short-rod-shaped halotolerant bacterium,designated CUG00002T,was isolated from the sediment of Xiaochaidan Lake of Qadam Basin,China,using R2A medium.The cells are Gram-negative,aerobic and forming creamy and circular colonies with diameter of 1-展开更多
In this work, the protein pattern of novel Halomonas smyrnensis AAD6T was compared to that of Halomonas salina DSMZ5928T, which is the closest species on the basis of 16S rRNA sequence, to understand how AAD6T differs...In this work, the protein pattern of novel Halomonas smyrnensis AAD6T was compared to that of Halomonas salina DSMZ5928T, which is the closest species on the basis of 16S rRNA sequence, to understand how AAD6T differs from type strains. Using high resolution NEPHEGE technique, the whole cell protein composition patterns of both Halomonas salina DSMZ5928T and H. smyrnensis AAD6T were mapped. The expressed proteins of the two microorganisms were mostly located at the acidic side of the gels, at molecular weight values of 60 to 17 kDa, and at isoelectric points 3.8 to 6.0, where they share a significant number of common protein spots. Identification and characterization of protein spots via whole genome sequencing data indicated that these two microorganisms used similar pathways, especially TCA cycle, for their survival;in other words, for their energy requirements. On the other hand, the protein expression differences in AAD6T and H. salina DSMZ 5928T showed that they prefer different metabolic pathways for lipid biosynthesis and in adaptation to extreme environments. Thus, we suggested that phylogenetic dissimilarities between these microorganisms could be related to the protein expression differences;in other words, metabolic flux differences in AAD6T and H. salina DSMZ 5928T. This is the first study to explain the dissimilarities of phenotypic characters and DNA-DNA hybridization between type strain and novel strain AAD6T by using protein expression differences.展开更多
With the rapid development of the marine economy,marine microbiologically influenced corrosion(MIC)has garnered increasing attention.However,most studies have not analyzed the MIC process over continuous and extended ...With the rapid development of the marine economy,marine microbiologically influenced corrosion(MIC)has garnered increasing attention.However,most studies have not analyzed the MIC process over continuous and extended periods,failing to provide a comprehensive understanding of MIC mechanisms at different stages.In this study,the corrosion behavior of EH36 steel caused by Halomonas titanicae in an aerobic enriched seawater over a 30-d incubation period was investigated driven by big data.The results revealed that the corrosion by H.titanicae against EH36 steel evolved dynamically over time.During the initial stages,the aerobic respiration of H.titanicae consumed significant amounts of oxygen,which suppressed the cathodic oxygen reduction process,thereby inhibiting corrosion compared to the abiotic conditions.As time progressed,the accumulation of corrosion products slowed the abiotic corrosion,while the biotic corrosion accelerated due to a shift from aerobic to anaerobic respiration by H.titanicae,utilizing Fe0 and nitrate as electron donors and acceptors,respectively.The big data results are consistent with the weight loss and electrochemical data,demonstrating the reliability of using big data monitoring techniques to characterize microbial corrosion processes.展开更多
Halomonas bluephagenesis TD serves as an exceptional chassis for next generation industrial biotechnology to produce various products.However,the simultaneous editing of multiple loci in H.bluephagenesis TD remains a ...Halomonas bluephagenesis TD serves as an exceptional chassis for next generation industrial biotechnology to produce various products.However,the simultaneous editing of multiple loci in H.bluephagenesis TD remains a significant challenge.Herein,we report the development of a multiple loci genome editing system,named CRISPR-deaminase-assisted base editor(CRISPR-BE)in H.bluephagenesis TD.This system comprises two components:a cytidine(CRISPR-cBE)and an adenosine(CRISPR-aBE)deaminase-based base editor.CRISPR-cBE can introduce a cytidine to thymidine mutation with an efficiency of up to 100%within a 7-nt editing window in H.bluephagenesis TD.Similarly,CRISPR-aBE demonstrates an efficiency of up to 100%in converting adenosine to guanosine mutation within a 7-nt editing window.CRISPR-cBE has been further validated and successfully employed for simultaneous multiplexed editing in H.bluephagenesis TD.Our findings reveal that CRISPR-cBE efficiently inactivated all six copies of the IS1086 gene simultaneously by introducing stop codon.This system achieved an editing efficiency of 100%and 41.67%in inactivating two genes and three genes,respectively.By substituting the Pcas promoter with the inducible promoter PMmp1,we optimized CRISPR-cBE system and ultimately achieved 100%editing efficiency in inactivating three genes.In conclusion,our research offers a robust and efficient method for concurrently modifying multiple loci in H.bluephagenesis TD,opening up vast possibilities for industrial applications in the future.展开更多
Acetoin is an important platform chemical,which has a wide range of applications in many industries.Halomonas bluephagenesis,a chassis for next generation of industrial biotechnology,has advantages of fast growth and ...Acetoin is an important platform chemical,which has a wide range of applications in many industries.Halomonas bluephagenesis,a chassis for next generation of industrial biotechnology,has advantages of fast growth and high tolerance to organic acid salts and alkaline environment.Here,α-acetolactate synthase andα-acetolactate decarboxylase from Bacillus subtilis 168 were co-expressed in H.bluephagenesis to produce acetoin from pyruvate.After reaction condition optimization and further increase ofα-acetolactate decarboxylase expression,acetoin production and yield were significantly enhanced to 223.4 mmol·L^(-1) and 0.491 mol·mol^(-1) from 125.4 mmol·L^(-1) and 0.333 mol·mol^(-1),respectively.Finally,the highest titer of 974.3 mmol·L^(-1)(85.84 g·L^(-1))of acetoin was accumulated from 2143.4 mmol·L^(-1)(188.6 g·L^(-1))of pyruvic acid within 8 h in fed-batch bioconversion under optimal reaction conditions.Moreover,the reusability of the cell catalysis was also tested,and the result illustrated that the whole-cell catalysis obtained 433.3,440.2,379.0,442.8 and 339.4 mmol·L^(-1)(38.2,38.8,33.4,39.0 and 29.9 g·L^(-1))acetoin in five repeated cycles under the same conditions.This work therefore provided an efficient H.bluephagenesis whole-cell catalysis with a broad development prospect in biosynthesis of acetoin.展开更多
文摘对一株合成聚-β-羟基脂肪酸酯(PHB)的Halomonas DQ-4发酵过程中碳、碳氮比、温度、pH值、转速及装液量等条件因子进行了优化。结果表明,最佳发酵条件为:碳源为葡萄糖,碳氮比为100/7,生长温度38℃,pH值为9,转速为160 r·min^(-1),装液量为100 m L/500 m L,连续培养60 h时,PHB产量达到峰值,为5.88 g·L^(-1)。同时,对发酵合成的PHB进行了核磁共振氢谱、红外光谱的表征,热重分析表明,PHB具有良好的热稳定性,并且在降解实验中表现了优良的生物降解性。
基金supported financially by the National Natural Science Foundation of China(Nos.U1660118 and 51871050)the National Environmental Corrosion Platform(NECP)of Chinathe Fundamental Research Funds for the Central Universities of Ministry of Education of China(No.N180205021).
文摘Algae are reported to be corrosive,while little is known about the role of the algae associated bacteria in the corrosion process.In the present study,Halomonas titanicae was isolated from a culture of an alga strain,Spirulina platensis,and identified through 16 S rRNA gene analysis.Corrosion behavior of 304L stainless steel(SS)coupons in the presence and absence of H.titanicae was characterized by using electrochemical measurements and surface analysis.The results showed that H.titanicae significantly accelerated the corrosion rate and decreased the pitting potential of 304L SS in the biotic medium.After removal of the corrosion products and biofilms,severe pitting corrosion caused by H.titanicae was observed.The largest pit depth after 14 d reached 6.6μm,which was 5.5 times higher than that of the sterile control(1.2μm).This is the first report revealing that an alga associated bacterium can induce microbiologically influenced corrosion(MIC),and a further concern is raised that whether algae play a role in the MIC process.
基金Supported by the Science & Technology Project of Nantong(No.AS2011012)the National Key Technology Research and Development Program(No.2011BAE06B04-05)the Open Project Program of the Key Laboratory of Marine Bio-resources Sustainable Utilization,SCSIO,CAS(No.LMB121006)
文摘In recent years,red tides occurred frequently in coastal areas worldwide.Various methods based on the use of clay,copper sulfate,and bacteria have been successful in controlling red tides to some extent.As a new defensive agent,marine microorganisms are important sources of compounds with potent inhibitory bioactivities against red-tide microalgae,such as Gymnodinium sp.(Pyrrophyta).In this study,we isolated a marine bacterium,HSB07,from seawater collected from Hongsha Bay,Sanya,South China Sea.Based on its 16S rRNA gene sequence and biochemical characteristics,the isolated strain HSB07 was identified as a member of the genus Halomonas.A crude ethyl acetate extract of strain HSB07 showed moderate inhibition activity against Gymnodinium sp.in a bioactive prescreening experiment.The extract was further separated into fractions A,B,and C by silica gel column chromatography.Fractions B and C showed strong inhibition activities against Gymnodinium.This is the first report of inhibitory activity of secondary metabolites of a Halomonas bacterium against a red-tide-causing microalga.
基金The COMRA Program under contract No. DY135-B2-01the Xiamen Ocean Economic Innovation and Development Demonstration Project under contract No. 16PZP001SF16the National Infrastructure of Natural Resources for Science and Technology Program of China under contract No. NIMR-2017-9。
文摘Nitrification is a key step in the global nitrogen cycle.Compared with autotrophic nitrification,heterotrophic nitrification remains poorly understood.In this study,Halomonas venusta MA-ZP17-13,isolated from seawater in shrimp aquaculture (Penaeus vannamei),could simultaneously undertake nitrification and denitrification.With the initial ammonium concentration at 100 mg/L,the maximum ammonium-nitrogen removal rate reached98.7%under the optimal conditions including C/N concentration ratio at 5.95,p H at 8.93,and Na Cl at 2.33%.The corresponding average removal rate was 1.37 mg/(L·h)(according to nitrogen) in 3 d at 11.2℃.By whole genome sequencing and analysis,nitrification-and denitrification-related genes were identified,including ammonia monooxygenase,nitrate reductase,nitrite reductase,nitric oxide dioxygenase and nitric oxide synthase;while no gene encoding hydroxylamine oxidase was identified,it implied the existence of a novel nitrification pathway from hydroxylamine to nitrate.These results indicate heterotrophic bacterium H.venusta MA-ZP17-13 can undertake simultaneous nitrification and denitrification at low temperature and has potential for NH_(4)^(+)-N/NH_(3)-N removal in marine aquaculture systems.
基金Supported by the National Natural Science Foundation of China(Nos.51308245,31570061)the Open Project of Jiangsu Provincial Engineering Laboratory for Advanced Materials of Salt Chemical Industry(No.SF201408)the Open Project of Jiangsu Provincial Key Construction Laboratory of Probiotics Preparation(No.JSYSZJ2017006)
文摘Halomonas sp.YSR-3 was isolated from the Yellow Sea and identified as a polyphosphate-accumulating bacterium and the characteristics of its intracellular polyphosphate(polyP)granules and phosphorus absorption were studied.Most YSR-3 cells stored one or two polyP granules in regular appearance and high-density.The diameter of the granules was about 400 nm measuring by a transmission electron microscope(TEM).After stained with 4,6-diamidino-2-phenylindole(DAPI)and visualized by a fluorescence microscope,the cells turned blue and the granules were bright yellow.The composition of granules includes P(major ingredient),Mg,S,K,and Ca as detected by an energy dispersive X-ray spectrometer(EDS).When inorganic phosphorus(po34-)and ferric ion(Fe3+)were added into media,the biomass increased and the cells formed intracellular polyP granules owing to the phosphorus assimilation from media.The YSR-3 obtained higher biomass by adding 0.02 g/L FePO4 than 0.005 g/L and 0.01 g/L FePO4;however,the phosphorus absorption was higher with 0.01 g/L FePO4 than 0.005 g/L and 0.02 g/L FePO4.The optical density at wavelength 480 nm(OD480nm)was 0.79 and 100%cells could form intracellular polyP granules.These results show that strain YSR-3 is able to acquire higher biomass and absorb more inorganic phosphorus when 0.01 g/L FePO4 is added.The characteristics of absorbing and storing phosphorus as intracellular inorganic polyP granules have a potential for application in high-efficiency phosphorus removal in wastewater treatment.
基金Supported by the National Basic Research Program of China (973 Program) (No. 2006AA10A401)the National Natural Science Foundation of China (No. 40376048)
文摘Hydantoinase is involved in the production of optically pure amino acids from racemic 5-mono-substituted hydantoins. We measured the D-hydantoinase activity in marine Halomonas sp. YSR-3 and amplified the D-hydantoinase gene by PCR. The gene was inserted into vector pGM-T and transformed into E. coli TOP10. The positive transformants with the D-hydantoinase gene were sequenced. The sequenced fragment comprises 1 510 base pairs. The D-hydantoinase gene from YSR-3 is 77% similar to that from Pseudomonas entornophila L4 by searching against the NCBI databse. The protein product of the YSR-3 D-hydantoinase gene is 75%, 73%, and 70% similar to those from Pseudomonasfluorescens Pf-5, Marinornonas sp. MED121, and Burkholderia vietnamiensis G4, respectively. The difference of the D-hydantoinase gene between marine Halornonas sp. YSR-3 and other terrestrial organisms is distinct.
基金supported by the National Natural Science Foundation of China (Grant Nos. 41002123 & 41030211)Key Laboratory of Salt Lake Resources and Chemistry, Qinghai Institute of Salt Lake, Chinese Academy of Sciences (No. KLSLRC-KF-13-DX-5)+1 种基金State Key Laboratory of Biogeology and Environmental Geology, China University of Geosciences (No. GBL11201)the Fundamental Research Funds for National University, China University of Geosciences (Wuhan)
文摘A short-rod-shaped halotolerant bacterium,designated CUG00002T,was isolated from the sediment of Xiaochaidan Lake of Qadam Basin,China,using R2A medium.The cells are Gram-negative,aerobic and forming creamy and circular colonies with diameter of 1-
文摘In this work, the protein pattern of novel Halomonas smyrnensis AAD6T was compared to that of Halomonas salina DSMZ5928T, which is the closest species on the basis of 16S rRNA sequence, to understand how AAD6T differs from type strains. Using high resolution NEPHEGE technique, the whole cell protein composition patterns of both Halomonas salina DSMZ5928T and H. smyrnensis AAD6T were mapped. The expressed proteins of the two microorganisms were mostly located at the acidic side of the gels, at molecular weight values of 60 to 17 kDa, and at isoelectric points 3.8 to 6.0, where they share a significant number of common protein spots. Identification and characterization of protein spots via whole genome sequencing data indicated that these two microorganisms used similar pathways, especially TCA cycle, for their survival;in other words, for their energy requirements. On the other hand, the protein expression differences in AAD6T and H. salina DSMZ 5928T showed that they prefer different metabolic pathways for lipid biosynthesis and in adaptation to extreme environments. Thus, we suggested that phylogenetic dissimilarities between these microorganisms could be related to the protein expression differences;in other words, metabolic flux differences in AAD6T and H. salina DSMZ 5928T. This is the first study to explain the dissimilarities of phenotypic characters and DNA-DNA hybridization between type strain and novel strain AAD6T by using protein expression differences.
基金financially supported by the National Natural Science Foundation of China(Nos.U2106206,52471079,42276212,and 42176043)the Natural Science Foundation of Shandong Province(ZR2024ME047)+1 种基金the National Materials Corrosion and Protection Data Center(No.2023DATAFU20-01)The authors wish to acknowledge Sen Wang,Haiyan Yu,Xiaomin Zhao from State Key Laboratory of Microbial Technology,Shandong University for the assistance in the SEM analysis。
文摘With the rapid development of the marine economy,marine microbiologically influenced corrosion(MIC)has garnered increasing attention.However,most studies have not analyzed the MIC process over continuous and extended periods,failing to provide a comprehensive understanding of MIC mechanisms at different stages.In this study,the corrosion behavior of EH36 steel caused by Halomonas titanicae in an aerobic enriched seawater over a 30-d incubation period was investigated driven by big data.The results revealed that the corrosion by H.titanicae against EH36 steel evolved dynamically over time.During the initial stages,the aerobic respiration of H.titanicae consumed significant amounts of oxygen,which suppressed the cathodic oxygen reduction process,thereby inhibiting corrosion compared to the abiotic conditions.As time progressed,the accumulation of corrosion products slowed the abiotic corrosion,while the biotic corrosion accelerated due to a shift from aerobic to anaerobic respiration by H.titanicae,utilizing Fe0 and nitrate as electron donors and acceptors,respectively.The big data results are consistent with the weight loss and electrochemical data,demonstrating the reliability of using big data monitoring techniques to characterize microbial corrosion processes.
基金supported by the National Natural Science Foundation of China(Grant No.32171415)the Chongqing Talents Top Youth Talent Program(No.CQYC202105065).
文摘Halomonas bluephagenesis TD serves as an exceptional chassis for next generation industrial biotechnology to produce various products.However,the simultaneous editing of multiple loci in H.bluephagenesis TD remains a significant challenge.Herein,we report the development of a multiple loci genome editing system,named CRISPR-deaminase-assisted base editor(CRISPR-BE)in H.bluephagenesis TD.This system comprises two components:a cytidine(CRISPR-cBE)and an adenosine(CRISPR-aBE)deaminase-based base editor.CRISPR-cBE can introduce a cytidine to thymidine mutation with an efficiency of up to 100%within a 7-nt editing window in H.bluephagenesis TD.Similarly,CRISPR-aBE demonstrates an efficiency of up to 100%in converting adenosine to guanosine mutation within a 7-nt editing window.CRISPR-cBE has been further validated and successfully employed for simultaneous multiplexed editing in H.bluephagenesis TD.Our findings reveal that CRISPR-cBE efficiently inactivated all six copies of the IS1086 gene simultaneously by introducing stop codon.This system achieved an editing efficiency of 100%and 41.67%in inactivating two genes and three genes,respectively.By substituting the Pcas promoter with the inducible promoter PMmp1,we optimized CRISPR-cBE system and ultimately achieved 100%editing efficiency in inactivating three genes.In conclusion,our research offers a robust and efficient method for concurrently modifying multiple loci in H.bluephagenesis TD,opening up vast possibilities for industrial applications in the future.
基金supported by the National Key Research and Development Program of China (Grant No.2018YFA0900200)the National Natural Science Foundation of China (Grant No.NSFC-21621004).
文摘Acetoin is an important platform chemical,which has a wide range of applications in many industries.Halomonas bluephagenesis,a chassis for next generation of industrial biotechnology,has advantages of fast growth and high tolerance to organic acid salts and alkaline environment.Here,α-acetolactate synthase andα-acetolactate decarboxylase from Bacillus subtilis 168 were co-expressed in H.bluephagenesis to produce acetoin from pyruvate.After reaction condition optimization and further increase ofα-acetolactate decarboxylase expression,acetoin production and yield were significantly enhanced to 223.4 mmol·L^(-1) and 0.491 mol·mol^(-1) from 125.4 mmol·L^(-1) and 0.333 mol·mol^(-1),respectively.Finally,the highest titer of 974.3 mmol·L^(-1)(85.84 g·L^(-1))of acetoin was accumulated from 2143.4 mmol·L^(-1)(188.6 g·L^(-1))of pyruvic acid within 8 h in fed-batch bioconversion under optimal reaction conditions.Moreover,the reusability of the cell catalysis was also tested,and the result illustrated that the whole-cell catalysis obtained 433.3,440.2,379.0,442.8 and 339.4 mmol·L^(-1)(38.2,38.8,33.4,39.0 and 29.9 g·L^(-1))acetoin in five repeated cycles under the same conditions.This work therefore provided an efficient H.bluephagenesis whole-cell catalysis with a broad development prospect in biosynthesis of acetoin.
