Metabolic Engineering Communications
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2018
·
IF 0
Xiaojing Jia,Robert M. Kelly,Yejun Han
( R )-acetoin is a four-carbon platform compound used as the precursor for synthesizing novel optically active materials. Ethylene glycol (EG) is a large-volume two-carbon commodity chemical used as the anti-freezing agent and building-block molecule for various polymers. Currently established microbial fermentation processes for converting monosaccharides to either ( R )-acetoin or EG are plagued by the formation of undesirable by-products. We show here that a cell-free bioreaction scheme can generate enantiomerically pure acetoin and EG as co-products from biomass-derived D -xylose. The seven-step, ATP-free system included in situ cofactor regeneration and recruited enzymes from Escherichia coli W3110, Bacillus subtilis shaijiu 32 and Caulobacter crescentus CB 2. Optimized in vitro biocatalytic conditions generated 3.2 mM ( R )-acetoin with stereoisomeric purity of 99.5% from 10 mM D -xylose at 30 °C and pH 7.5 after 24 h, with an initial ( R )-acetoin productivity of 1.0 mM/h. Concomitantly, EG was produced at 5.5 mM, with an initial productivity of 1.7 mM/h. This in vitro biocatalytic platform illustrates the potential for production of multiple value-added biomolecules from biomass-based sugars with no ATP requirement.
研究领域:
生物化学
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合成生物学
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代谢工程
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DOI
PubMed
Energies
·
2022
·Vol. 15
·期 12
·4242
·
IF 3.252
Xue-Yong Li,Jia-Ying Huang,Qiang Zhou,Yuan-Yuan Xu,Ponnandy Prabhu,Ye-Wang Zhang
Acetoin, a four-carbon hydroxyl-keto compound, is used in the food, pharmaceutical, and chemical industries. The cascade enzymatic production is considered a promising and efficient method to produce acetoin. However, the stability and compatibility of the enzymes under the same catalytic conditions are challenges that need to be resolved. In this work, alcohol dehydrogenase, acetaldehyde lyase, and NADH oxidase were selected to work at the same conditions to efficiently convert ethanol into acetoin. These three enzymes were immobilized on epoxy-modified magnetic nanomaterials to obtain highly stable biocatalysts. The stability and the immobilization conditions, including temperature, pH, enzyme–carrier ratio, and immobilization time, were optimized to obtain the immobilized enzymes with a high catalytic activity. The cascade reactions catalyzed by the immobilized enzymes yielded a high conversion of 90%, suggesting that the use of immobilized enzymes is a promising way to produce acetoin.Keywords:immobilization;NADH oxidase;acetaldehyde lyase;alcohol dehydrogenase;acetoin;cascade enzymatic reactions
研究领域:
生物技术
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酶学
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化学工程
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产品关联
DOI
高影响因子
LWT-FOOD SCIENCE AND TECHNOLOGY
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2025
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IF 6.6
Yixuan LUO,Huiting LAI,Shuhua LIN,Qi TANG,Juhua ZHANG,Wei LIU
Acetoin (ACT) is an important flavor compound found in traditional fermented foods, and a precursor of compounds such as pyrazine, which is a direct source of soy sauce aroma in fermented soybean products. In this study, four acetoinproducing lactic acid bacteria (LAB) were screened from traditional high-salt fermented foods in Hunan Province, China, by the creatine colorimetric method, Gram staining, and catalase test. QT038 and JC094 exhibited the highest acetoin contents of 81.3 mg/L and 76.8 mg/L. JC094, QT019, and LJ064 were determined to be Lactiplantibacillus plantarum , and QT038 was determined to be Lactiplantibacillus paraplantarum . The growth characteristics, temperature tolerance, salt tolerance, and antioxidant capacity of the 4 strains were studied. The strains QT038 and JC094 showed strong tolerance and antioxidant characteristics. Volatile analysis of fermented soy sauce simulation experiment with selected strains revealed JC094 fermented samples contained significantly more flavor compounds, both in variety (38) and total content (3.57 mg/L), compared to other strains. Notably, QT038 and JC094 improved the acetoin levels (0.161 mg/L). The results demonstrated that QT038 and JC094 have the ability to produce higher levels of acetoin to enhance flavor quality and can be further mixed or co-fermented with yeasts in soy sauce fermentation.
研究领域:
食品科学
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微生物学
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风味化学
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生物技术
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产品关联
DOI
高影响因子
COMPUTERS IN BIOLOGY AND MEDICINE
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2023
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IF 7.7
Jinyi Qian,Yuzhou Wang,Xiner Liu,Zijian Hu,Nan Xu,Yuetong Wang,Tianqiong Shi,Chao Ye
Acetoin was widely used in food, medicine, and other industries, because of its unique fragrance. Bacillus amyloliquefaciens was recognized as a safe strain and a promising acetoin producer in fermentation. However, due to the complexity of its metabolic network, it had not been fully utilized. Therefore, a genome-scale metabolic network model ( i JYQ746) of B. amyloliquefaciens was constructed in this study, containing 746 genes, 1736 reactions, and 1611 metabolites. The results showed that Mg 2+ , Mn 2+ , and Fe 2+ have inhibitory effects on acetoin. When the stirring speed was 400 rpm, the maximum titer was 49.8 g L −1 . Minimization of metabolic adjustments (MOMA) was used to identify potential metabolic modification targets 2-oxoglutarate aminotransferase ( serC , EC 2.6.1.52) and glucose-6-phosphate isomerase ( pgi , EC 5.3.1.9). These targets could effectively accumulate acetoin by increasing pyruvate content, and the acetoin synthesis rate was increased by 610% and 10%, respectively. This provides a theoretical basis for metabolic engineering to reasonably transform B. amyloliquefaciens and produce acetoin.
研究领域:
生物技术
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代谢工程
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微生物学
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产品关联
DOI
PubMed
高影响因子
Biocatalysis and Agricultural Biotechnology
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2019
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IF 0
Feng-Qin Gong,Qi-Shun Liu,Hai-Dong Tan,Tang Li,Cheng-Yu Tan,Heng Yin
Acetoin (3-hydroxy-2-butanone) is an important four-carbon compound widely used in the food industry and other industrial applications. This study aimed to identify a novel butanediol dehydrogenase that can efficiently catalyze the formation of acetoin. A novel butanediol dehydrogenase, Bt BDH, was identified from Bacillus thuringiensis subsp. Kurstaki ACCC 10066, and its enzymatic properties were characterized. The optimum pH and temperature for the oxidation activity of Bt BDH were 10.0 and 50 °C, respectively, and those for the reduction activities were 7.5 and 35 °C, respectively. In addition, it exhibits stability over a wide pH range (6–10) and temperatures up to 70 °C. Bt BDH showed good stability after storage for 3 months at 4 °C. Moreover, ethylenediaminetetracetic acid (EDTA) inhibits the enzymatic activity of Bt BDH, indicating that the enzyme is metal-dependent. This study characterized a novel (2R,3R) −2,3-butanediol dehydrogenase. Its excellent oxidation activity and stability ensure its great industrial application potential in the production of acetoin.
研究领域:
生物化学
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微生物学
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酶学
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产品关联
DOI
NUCLEIC ACIDS RESEARCH
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2023
·Vol. 51
·期 21
·11952-11966
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IF 14.9
He Hehe,Li Youran,Ma Xufan,Xu Sha,Zhang Liang,Ding Zhongyang,Shi Guiyang
Synthetic regulation of metabolic fluxes has emerged as a common strategy to improve the performance of microbial cell factories. The present regulatory toolboxes predominantly rely on the control and manipulation of carbon pathways. Nitrogen is an essential nutrient that plays a vital role in growth and metabolism. However, the availability of broadly applicable tools based on nitrogen pathways for metabolic regulation remains limited. In this work, we present a novel regulatory system that harnesses signals associated with nitrogen metabolism to redirect excess carbon flux in Bacillus licheniformis. By engineering the native transcription factor GlnR and incorporating a sorbitol-responsive element, we achieved a remarkable 99% inhibition of the expression of the green fluorescent protein reporter gene. Leveraging this system, we identified the optimal redirection point for the overflow carbon flux, resulting in a substantial 79.5% reduction in acetoin accumulation and a 2.6-fold increase in acetate production. This work highlight the significance of nitrogen metabolism in synthetic biology and its valuable contribution to metabolic engineering. Furthermore, our work paves the way for multidimensional metabolic regulation in future synthetic biology endeavors.
研究领域:
合成生物学
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代谢工程
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微生物生物技术
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DOI
PubMed
高影响因子
RSC Advances
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2018
·Vol. 8
·期 53
·30512-30519
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IF 2.936
Jian-Xiu Li,Yan-Yan Huang,Xian-Rui Chen,Qi-Shi Du,Jian-Zong Meng,Neng-Zhong Xie,Ri-Bo Huang
Acetoin is an important platform chemical with a variety of applications in foods, cosmetics, chemical synthesis, and especially in the asymmetric synthesis of optically active pharmaceuticals. It is also a useful breath biomarker for early lung cancer diagnosis. In order to enhance production of optical (S)-acetoin and facilitate this building block for a series of chiral pharmaceuticals derivatives, we have developed a systematic approach using in situ-NADH regeneration systems and promising diacetyl reductase. Under optimal conditions, we have obtained 52.9 g L−1 of (S)-acetoin with an enantiomeric purity of 99.5% and a productivity of 6.2 g (L h)−1. The results reported in this study demonstrated that the production of (S)-acetoin could be effectively improved through the engineering of cofactor regeneration with promising diacetyl reductase. The systematic approach developed in this study could also be applied to synthesize other optically active α-hydroxy ketones, which may provide valuable benefits for the study of drug development.
研究领域:
生物催化
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药物化学
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代谢工程
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生物技术
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产品关联
DOI
PubMed
高影响因子
Scientific Reports
·
2017
·Vol. 7
·期 1
·1-10
·
IF 4.259
Jia Xiaojing,Liu Ying,Han Yejun
Acetoin (3-hydroxy-2-butanone) is an important bio-based platform chemical with wide applications. In vitro enzyme catalysed synthesis exhibits great feasibility in the production of chemicals with high purity. In the present work, a synthetic pathway involving a two-step continuous reaction was constructed in vitro for acetoin production from pyruvate at improved temperature. Thermostable candidates, acetolactate synthase (coAHASL1 and coAHASL2 from Caldicellulosiruptor owensensis OL) and α-acetolactate decarboxylase (bsALDC from Bacillus subtilis IPE5-4) were cloned, heterologously expressed, and characterized. All the enzymes showed maximum activities at 65–70 °C and pH of 6.5. Enzyme kinetics analysis showed that coAHASL1 had a higher activity but lower affinity against pyruvate than that of coAHASL2. In addition, the activities of coAHASL1 and bsALDC were promoted by Mn 2+ and NADPH. The cascade enzymatic reaction was optimized by using coAHASL1 and bsALDC based on their kinetic properties. Under optimal conditions, a maximum concentration of 3.36 ± 0.26 mM acetoin was produced from 10 mM pyruvate after reaction for 24 h at 65 °C. The productivity of acetoin was 0.14 mM h −1 , and the yield was 67.80% compared with the theoretical value. The results confirmed the feasibility of synthesis of acetoin from pyruvate with a cell-free enzyme catalysed system at improved temperature.
研究领域:
生物化学
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酶学
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生物技术
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产品关联
DOI
PubMed
高影响因子
Frontiers in Bioengineering and Biotechnology
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2022
·
IF 6.064
Jing Li,Jian Lu,Zhilin Ma,Jianxiu Li,Xianrui Chen,Mengxue Diao,Nengzhong Xie
2,3,5,6-Tetramethylpyrazine (TMP) is an active pharmaceutical ingredient originally isolated from Ligusticum wallichii for curing cardiovascular and cerebrovascular diseases and is widely used as a popular flavoring additive in the food industry. Hence, there is a great interest in developing new strategies to produce this high-value compound in an ecological and economical way. Herein, a cost-competitive combinational approach was proposed to accomplish green and high-efficiency production of TMP. First, microbial cell factories were constructed to produce acetoin (3-hydroxy-2-butanone, AC), an endogenous precursor of TMP, by introducing a biosynthesis pathway coupled with an intracellular NAD + regeneration system to the wild-type Escherichia coli . To further improve the production of ( R )-AC, the metabolic pathways of by-products were impaired or blocked stepwise by gene manipulation, resulting in 40.84 g/L ( R )-AC with a high optical purity of 99.42% in shake flasks. Thereafter, an optimal strain designated GXASR11 was used to convert the hydrolysates of inexpensive feedstocks into ( R )-AC and achieved a titer of 86.04 g/L within 48 h in a 5-L fermenter under optimized fermentation conditions. To the best of our knowledge, this is the highest ( R )-AC production with high optical purity (≥98%) produced from non-food raw materials using recombinant E . coli . The supernatant of fermentation broth was mixed with diammonium phosphate (DAP) to make a total volume of 20 ml and transferred to a high-pressure microreactor. Finally, 56.72 g/L TMP was obtained in 3 h via the condensation reaction with a high conversion rate (85.30%) under optimal reaction conditions. These results demonstrated a green and sustainable approach to efficiently produce high-valued TMP, which realized value addition of low-cost renewables.
研究领域:
药学
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生物技术
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代谢工程
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可持续化学
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微生物学
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产品关联
DOI
PubMed
高影响因子
INDUSTRIAL CROPS AND PRODUCTS
·
2025
·
IF 6.2
Ziqian Gao,Ning Zhu,Minhui Yang,Xingshuai Cui,Changze Han,Xiaopeng Guo,Feifan Leng,Ziyi Cheng,Wen Luo,Yonggang Wang
This study aims to explore the mechanisms underlying the interactions between the secondary metabolism and root-associated bacteria of Codonopsis pilosula . This study collected root and rhizosphere soil samples from various C. pilosula cultivars and employed metabolome, transcriptome, and microbiome analysis to assess the variations in volatile organic compounds (VOCs), root-associated bacterial communities, terpenoid biosynthetic genes, and terpenoid metabolites across these cultivars. Significant differences were observed in VOC secretion from roots, terpenoid metabolite accumulation, and the composition and structure of root-associated bacterial communities among the cultivars. A total of 57 differentially expressed genes within the terpenoid synthesis pathway were identified, and terpenoid biosynthetic pathways in C. pilosula were constructed by integrating terpenoid differential metabolites. Furthermore, correlation analysis between VOCs and root-associated bacteria suggested that certain VOCs, such as acetoin, may play a role in shaping the composition of the root microbiome, particularly with respect to bacteria like Nitrospira and RB41 . Acetoin had been experimentally proven to promote the germination of C. pilosula seeds. Additionally, correlations between root-associated bacteria and both terpenoid synthesis genes and metabolites suggested that bacteria such as Nitrospira may influence the synthesis of terpenoid compounds like abscisic acid by modulating the expression of related biosynthetic genes. Collectively, this study provides preliminary insights into the mechanisms of secondary metabolite-root-associated bacterial interactions in C. pilosula , offering valuable guidance for the scientific cultivation and enhancement of the medicinal value of C. pilosula in future.
研究领域:
植物科学
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微生物学
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代谢组学
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转录组学
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产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2025
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IF 7.7
Xudong Liang,Ertai Na,Guowen Zhong,Nuo Yang,Jiamin Chen,Jiayu Li,Xiaozhan Yang,Ruiqin Gao,Wenlin Feng,Guo-Dong Li
The foodborne pathogen Listeria monocytogenes poses a severe threat to public health due to its high virulence and environmental adaptability, necessitating the development of sensing technologies for real-time monitoring of early contamination. This investigation addresses the detection bottleneck of its specific metabolic marker, acetoin, by innovatively developing a citric acid-assisted sol-gel method that successfully synthesizes Cd 2 GeO 4 nanoparticles. This approach overcomes the critical challenge of controlling the nanostructure and surface defect states of Ge-based materials, which is often problematic with traditional high-temperature solid-state or hydrothermal methods. The experiments demonstrate that the Cd 2 GeO 4 sensor exhibits outstanding performance for acetoin. DFT calculations elucidated that the Cd-O sites on the (110) crystal plane achieve specific recognition through the chemisorption of the C O and OH groups of acetoin. These insights elucidate the mechanism behind acetoin sensing, suggesting that Cd 2 GeO 4 may be employed as a novel sensing material for Listeria monocytogenes detection.
研究领域:
纳米材料
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食品安全
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生物传感器
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病原体检测
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材料科学
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分析化学
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产品关联
DOI
高影响因子
Metabolomics
·
2019
·Vol. 15
·期 3
·1-12
·
IF 3.167
Yuan Honglun,Xu Yong,Chen Yaozhong,Zhan Yangyang,Wei Xuetuan,Li Lu,Wang Dong,He Penghui,Li Shengqing,Chen Shouwen
Introduction Acetoin serves as a high value-added platform with a broad range of applications, and can be effectively produced by Bacillus licheniformis . However, its toxicity to the producing strain hinders the higher acetoin production, and current knowledge about the acetoin resistance mechanisms of B. licheniformis is quite limited. Objectives To comprehensively investigate the metabolic changes in B. licheniformis under acetoin stress. Methods We used gas chromatography-mass spectrometry based untargeted metabolomics approach to measure the metabolic profiles of B. licheniformis under 20, 40 and 80 g/L acetoin stress. Transcriptional analysis was conducted to verify the metabolomics results. Results A total of 119 metabolites were identified in our experiment. The metabolic responses of B. licheniformis to acetoin stress were as follows: (i) pentose phosphate pathway and tricarboxylic acid (TCA) cycle were negatively affected by acetoin stress. In turn, glyoxylate cycle was activated to supply malic acid. (ii) Acetoin stress induced the accumulation of serine, valine, leucine and protective osmolytes (glycine and proline). (iii) Acetoin stress induced a higher saturated fatty acid ratio, which indicated a lower fluidity of cell membrane that could inhibit the entry of acetoin into cytoplasm. (iv) Synthesis of phosphatidylserine was enhanced, and phosphatidylethanolamine content was probably increased under acetoin stress. Conclusions This study revealed the metabolic perturbations of B. licheniformis to acetoin stress. In response to acetoin stress, glyoxylate cycle was activated, protective osmolytes were accumulated, saturated fatty acid ratio was elevated and synthesis of phosphatidylserine was enhanced in B. licheniformis .
研究领域:
微生物学
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代谢组学
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生物技术
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）