LPS-Induced Acute Kidney Injury Is Mediated by Nox4-SH3YL1.

Cytosolic proteins are required for regulation of NADPH (nicotinamide adenine dinucleotide phosphate) oxidase (Nox) isozymes. Here we show that Src homology 3 (SH3) domain-containing YSC84-like 1 (SH3YL1), as a Nox4 cytosolic regulator, mediates lipopolysaccharide (LPS)-induced H2O2 generation, leading to acute kidney injury. The SH3YL1, Ysc84p/Lsb4p, Lsb3p, and plant FYVE proteins (SYLF) region and SH3 domain of SH3YL1 contribute to formation of a complex with Nox4-p22phox. Interaction of p22phox with SH3YL1 is triggered by LPS, and the complex induces H2O2 generation and pro-inflammatory cytokine expression in mouse tubular epithelial cells. After LPS injection, SH3YL1 knockout mice show lower levels of acute kidney injury biomarkers, decreased secretion of pro-inflammatory cytokines, decreased infiltration of macrophages, and reduced tubular damage compared with wild-type (WT) mice. The results strongly suggest that SH3YL1 is involved in renal failure in LPS-induced acute kidney injury (AKI) mice. We demonstrate that formation of a ternary complex of p22phox-SH3YL1-Nox4, leading to H2O2 generation, induces severe renal failure in the LPS-induced AKI model.

Reliability of Point-of-Care Hematocrit Measurement During Liver Transplantation.

BACKGROUND: Although point-of-care (POC) analyzers are commonly used during liver transplantation (LT), the accuracy of hematocrit measurement using a POC analyzer has not been evaluated. In this retrospective observational study, we aimed to evaluate the accuracy of hematocrit measurement using a POC analyzer and identify potential contributors to the measurement error and their influence on mistransfusion during LT. METHODS: We retrospectively collected 6461 pairs of simultaneous intraoperative hematocrit measurements using POC analyzers and laboratory devices during LTs in 901 patients. The agreement of hematocrit measurements was assessed using Bland-Altman analysis for repeated measurements, while the incidence and magnitude of hematocrit measurement error were compared among 16 different laboratory abnormality categories. A generalized estimating equation analysis was performed to identify potential contributors to falsely low-measured POC hematocrit. Additionally, we defined potential "overtransfusion" in the case when POC hematocrit was <20% and laboratory hematocrit was ≥20% and investigated its association with intraoperative transfusion. RESULTS: The POC hematocrit measurements were falsely lower than the laboratory hematocrit measurements in 70.3% (4541/6461) of pairs. The median (interquartile range) of hematocrit measurement error was -1.20 (-2.60 to 0.20). Bland-Altman analysis showed that 24.5% (1583/6461) of the errors were outside our a priori defined clinically acceptable limits of ±3%. The incidence of falsely low-measured hematocrit was significantly higher with the presence of concomitant hypoalbuminemia and hypoproteinemia. Hypoalbuminemia combined with hyperglycemia showed significantly larger hematocrit measurement error. Hypoalbuminemia, hypoproteinemia, and hyperglycemia were predictors of falsely low-measured hematocrit. Furthermore, the overtransfusion group showed larger amount of transfusion than the adequately transfused group, with a median difference of 2 units (95% confidence interval [0-4], P = .039), despite similar amount of blood loss. CONCLUSIONS: Hematocrit measured using the POC device tends to be lower than the laboratory hematocrit measured during LT. Commonly encountered laboratory abnormalities during LT include hypoalbuminemia, hypoproteinemia, and hyperglycemia, which may contribute to falsely low-measured POC hematocrit. Careful consideration of these confounders may help reduce overtransfusion that occurs due to falsely low-measured POC hematocrit.

Effect of head position on laryngeal visualisation with the McGrath MAC videolaryngoscope in paediatric patients: A randomised controlled trial.

BACKGROUND: The McGrath MAC video laryngoscope can improve visualisation of the glottis compared with the Macintosh direct laryngoscope. However, good visualisation of the glottis does not guarantee rapid or successful intubation because of difficulty in handling the McGrath device. OBJECTIVE: We evaluated the effect of head elevation, aligning the positions of the external auditory meatus and sternal notch in the horizontal plane, on visualisation of the glottis and handling of the McGrath laryngoscope in paediatric patients. DESIGN: A randomised controlled trial. SETTING: The operating rooms of our tertiary care hospital. PATIENTS: Forty-six children, American Society of Anaesthesiologists' physical status 1 or 2, aged 3 to 7 years. INTERVENTION: Videolaryngoscopy using the McGrath device was performed with the head either flat or elevated. MAIN OUTCOME MEASURES: The percentage of glottis opening score, the use of optimisation manoeuvre and time to successful tracheal intubation were recorded. RESULTS: The median (IQR) percentage of glottis opening score was higher after head elevation than when the head was flat in all patients [100 (100 to 100)% vs. 100 (90 to 100)%, P = 0.0001). The need for use of optimisation procedures (50 vs. 9%, P = 0.004) and mean (SD) time to intubation (17 ±â€Š4 s vs. 15 ±â€Š3 s, P = 0.008) were lower in the head-elevated group. CONCLUSION: Visualisation of the glottis and handling of the McGrath MAC video laryngoscope were significantly better when the external auditory meatus and sternal notch were aligned in the horizontal plane. TRIAL REGISTRATION: http://cris.nih.go.kridentifier:KCT0001443.

Deletion of the budBAC operon in Klebsiella pneumoniae to understand the physiological role of 2,3-butanediol biosynthesis.

Klebsiella pneumoniae is known to produce 2,3-butanediol (2,3-BDO), a valuable chemical. In K. pneumoniae, the 2,3-BDO operon (budBAC) is involved in the production of 2,3-BDO. To observe the physiological role of the 2,3-BDO operon in a mixed acid fermentation, we constructed a budBAC-deleted strain (SGSB109). The production of extracellular metabolites, CO2 emission, carbon distribution, and NADH/NAD(+) balance of SGSB109 were compared with the parent strain (SGSB100). When comparing the carbon distribution at 15 hr, four significant differences were observed: in 2,3-BDO biosynthesis, lactate and acetate production (lactate and acetate production increased 2.3-fold and 4.1-fold in SGSB109 compared to SGSB100), CO2 emission (higher in SGSB100), and carbon substrate uptake (higher in SGSB100). Previous studies on the inactivation of the 2,3-BDO operon were focused on the increase of 1,3-propanediol production. Few studies have been done observing the role of 2,3-BDO biosynthesis. This study provides a prime insight into the role of 2,3-BDO biosynthesis of K. pneumoniae.

Inactivation of the virulence factors from 2,3-butanediol-producing Klebsiella pneumoniae.

The microbiological production of 2,3-butanediol (2,3-BDO) has attracted considerable attention as an alternative way to produce high-value chemicals from renewable sources. Among the number of 2,3-BDO-producing microorganisms, Klebsiella pneumoniae has been studied most extensively and is known to produce large quantity of 2,3-BDO from a range of substrates. On the other hand, the pathogenic characteristics of the bacteria have limited its industrial applications. In this study, two major virulence traits, outer core LPS and fimbriae, were removed through homologous recombination from 2,3-BDO-producing K. pneumoniae 2242 to expand its uses to the industrial scale. The K. pneumoniae 2242 ∆wabG mutant strain was found to have an impaired capsule, which significantly reduced its ability to bind to the mucous layer and evade the phagocytic activity of macrophage. The association with the human ileocecal epithelial cell, HCT-8, and the bladder epithelial cell, T-24, was also reduced dramatically in the K. pneumoniae 2242 ∆fimA mutant strain that was devoid of fimbriae. However, the growth rate and production yield for 2,3-BDO were unaffected. The K. pneumoniae strains developed in this study, which are devoid of the major virulence factors, have a high potential for the efficient and sustainable production of 2,3-BDO.

Comparative whole genome transcriptome and metabolome analyses of five Klebsiella pneumonia strains.

The integration of transcriptomics and metabolomics can provide precise information on gene-to-metabolite networks for identifying the function of novel genes. The goal of this study was to identify novel gene functions involved in 2,3-butanediol (2,3-BDO) biosynthesis by a comprehensive analysis of the transcriptome and metabolome of five mutated Klebsiella pneumonia strains (∆wabG = SGSB100, ∆wabG∆budA = SGSB106, ∆wabG∆budB = SGSB107, ∆wabG∆budC = SGSB108, ∆wabG∆budABC = SGSB109). First, the transcriptomes of all five mutants were analyzed and the genes exhibiting reproducible changes in expression were determined. The transcriptome was well conserved among the five strains, and differences in gene expression occurred mainly in genes coding for 2,3-BDO biosynthesis (budA, budB, and budC) and the genes involved in the degradation of reactive oxygen, biosynthesis and transport of arginine, cysteine biosynthesis, sulfur metabolism, oxidoreductase reaction, and formate dehydrogenase reaction. Second, differences in the metabolome (estimated by carbon distribution, CO2 emission, and redox balance) among the five mutant strains due to gene alteration of the 2,3-BDO operon were detected. The functional genomics approach integrating metabolomics and transcriptomics in K. Pneumonia presented here provides an innovative means of identifying novel gene functions involved in 2,3-BDO biosynthesis metabolism and whole cell metabolism.

Industrial Production of 2,3-Butanediol from the Engineered Corynebacterium glutamicum.

The platform chemical 2,3-butanediol (2,3-BDO) is a valuable product that can be converted into several petroleum-based chemicals via simple chemical reactions. Here, we produced 2,3-BDO with the non-pathogenic and rapidly growing Corynebacterium glutamicum. To enhance the 2,3-BDO production capacity of C. glutamicum, we introduced budA encoding acetolactate decarboxylase from Klebsiella pneumoniae, a powerful 2,3-BDO producer. Additionally, budB (encoding α-acetolactate synthase) and budC (encoding acetoin reductase) were introduced from K. pneumoniae to reinforce the carbon flux in the 2,3-BDO production. Because budC had a negative effect on 2,3-BDO production in C. glutamicum, the budB and budA introduced strain, SGSC102, was selected for 2,3-BDO production, and batch culture was performed at 30 °C, 250 rpm and pH 6.86 with pure glucose, molasses, and cassava powder as carbon substrates. After batch culture, significant amount of 2,3-BDO (18.9 and 12.0 g/L, respectively) was produced from 80 g/L of pure glucose and cassava powder.

A non-pathogenic and optically high concentrated (R,R)-2,3-butanediol biosynthesizing Klebsiella strain.

The objective of this work was to construct a non-pathogenic Klebsiella pneumonia strain that can produce optically high concentrated (R,R)-2,3-BDO. A K. pneumonia mutant lacking the pathogenic factor was used as the host strain. In order to construct a K. pneumonia strain that would biosynthesize high concentrated (R,R)-2,3-BDO, gene deletion and over-expression methods were combined; firstly, the 2,3-BDO dehydrogenase (budC) gene was deleted to re-direct utilization of the carbon source to (R,R)-2,3-BDO biosynthesis; secondly, the two glycerol dehydrogenase (GDH) enzymes in K. pneumonia (DhaD and GldA) were over-expressed to maximize (R,R)-2,3-BDO biosynthesis; and thirdly, the lactate dehydrogenase (ldhA) gene was deleted to minimize the accumulation of lactate. SGSB112, a non-pathogenic strain of K. pneumonia that can produce optically high concentrated (R,R)-2,3-BDO, was constructed as above. Approximately 36% of the carbon source was converted to (R,R)-2,3-BDO by SGSB112, achieving a production of 61gL(-1) (R,R)-2,3-BDO in a fed-batch fermentation. On the other hand, meso-2,3-BDO was produced 1.4gL(-1) and (S,S)-2,3-BDO was not detected. This study provides an insight into 2,3-BDO biosynthesis in K. pneumonia and demonstrates the achievement of high-yield production of optically high concentrated (R,R)-2,3-BDO through constructing a strain by genetic modification and metabolic engineering.

The influence of budA deletion on glucose metabolism related in 2,3-butanediol production by Klebsiella pneumoniae.

Klebsiella pneumoniae (K. pneumoniae), which is a promising microorganism for industrial bulk production of 2,3-butanediol (2,3-BDO), naturally converts glucose to 2,3-BDO. The 2,3-BDO biosynthesis from glucose is composed of three steps; α-acetolactate biosynthesis by α-acetolactate synthase (budB); acetoin biosynthesis by α-acetolactate decarboxylase (budA); and 2,3-BDO biosynthesis by acetoin reductase (budC). In an effort to understand the influence of blocked 2,3-BDO pathway on K. pneumoniae glucose metabolism by budA deletion, we constructed K. pneumoniaeΔwabGΔbudA (SGSB106). Carbon flux distribution analysis, transcriptome analysis and extracellular amino acid concentration analysis were carried out to understand the effects of the budA deletion, and K. pneumoniaeΔwabG (SGSB100) was used as a control strain. Approximately 50.3% decrease in CO2 emission; and approximately 3.8-fold increase in amino acid production was observed in SGSB106. In addition to, among the amino acids, valine production significantly increased, suggesting that the branched-chain amino acid biosynthesis (BACC) in SGSB106 was activated by deletion of budA. Furthermore, whole genome transcriptome analysis of SGSB106 and SGSB100, correlates with the results from carbon distribution and amino acids concentration analyses.

Redistribution of carbon flux toward 2,3-butanediol production in Klebsiella pneumoniae by metabolic engineering.

Klebsiella pneumoniae KCTC2242 has high potential in the production of a high-value chemical, 2,3-butanediol (2,3-BDO). However, accumulation of metabolites such as lactate during cell growth prevent large-scale production of 2,3-BDO. Consequently, we engineered K. pneumoniae to redistribute its carbon flux toward 2,3-BDO production. The ldhA gene deletion and gene overexpression (budA and budB) were conducted to block a pathway that competitively consumes reduced nicotinamide adenine dinucleotide and to redirect carbon flux toward 2,3-BDO biosynthesis, respectively. These steps allowed efficient glucose conversion to 2,3-BDO under slightly acidic conditions (pH 5.5). The engineered strain SGSB105 showed a 40% increase in 2,3-BDO production from glucose compared with that of the host strain, SGSB100. Genes closely related to 2,3-BDO biosynthesis were observed at the gene transcription level by cultivating the SGSB100, SGSB103, SGSB104, and SGSB105 strains under identical growth conditions. Transcription levels for budA, budB, and budC increased approximately 10% during the log phase of cell growth relative to that of SGSB100. Transcription levels of 2,3-BDO genes in SGSB105 remained high during the log and stationary phases. Thus, the carbon flux was redirected toward 2,3-BDO production. Data on batch culture and gene transcription provide insight into improving the metabolic network for 2,3-BDO biosynthesis for industrial applications.

Testosterone stimulates Duox1 activity through GPRC6A in skin keratinocytes.

Testosterone is an endocrine hormone with functions in reproductive organs, anabolic events, and skin homeostasis. We report here that GPRC6A serves as a sensor and mediator of the rapid action of testosterone in epidermal keratinocytes. The silencing of GPRC6A inhibited testosterone-induced intracellular calcium ([Ca(2+)]i) mobilization and H2O2 generation. These results indicated that a testosterone-GPRC6A complex is required for activation of Gq protein, IP3 generation, and [Ca(2+)]i mobilization, leading to Duox1 activation. H2O2 generation by testosterone stimulated the apoptosis of keratinocytes through the activation of caspase-3. The application of testosterone into three-dimensional skin equivalents increased the apoptosis of keratinocytes between the granular and stratified corneum layers. These results support an understanding of the molecular mechanism of testosterone-dependent apoptosis in which testosterone stimulates H2O2 generation through the activation of Duox1.

Increased 2,3-butanediol production by changing codon usages in Escherichia coli.

The natural microorganism Escherichia coli without modification is not suitable for the efficient production of 2,3-butanediol (2,3-BD) on an industrial scale because of its poor metabolic performance. Metabolic capacities of E. coli have been improved to produce 2,3-BD efficiently, the performance of which is possible for producing such a product. Codon optimization with the ribosome-binding site for the efficient production of target genes (budA and budC) was achieved by molecular engineering, which allowed the metabolic engineering to proceed to the next level. As a result, comparing the productivity in 26 H, where the amount of p18COR was 1.04 g/L and that of p18WTR was 0.41 g/L, represents an approximate 60.6% increase in the productivity of the p18WTR with codon optimization. In other words, p18COR was 2.54-fold greater than p18WTR in the production of 2,3-BD.

The regulation of 2,3-butanediol synthesis in Klebsiella pneumoniae as revealed by gene over-expressions and metabolic flux analysis.

A variety of microorganism species are able naturally to produce 2,3-butanediol (2,3-BDO), although only a few of them are suitable for consideration as having potential for mass production purposes. Klebsiella pneumoniae (K. pneumoniae) is one such strain which has been widely studied and used industrially to produce 2,3-BDO. In the central carbon metabolism of K. pneumoniae, the 2,3-BDO synthesis pathway is dominated by three essential enzymes, namely acetolactate decarboxylase, acetolactate synthase, and butanediol dehydrogenase, which are encoded by the budA, budB, and budC genes, respectively. The mechanisms of the three enzymes have been characterized with regard to their function and roles in 2,3-BDO synthesis and cell growth (Blomqvist et al. in J Bacteriol 175(5):1392-1404, 1993), while a few studies have focused on the cooperative mechanisms of the three enzymes and their mutual interactions. Therefore, the K. pneumoniae KCTC2242::ΔwabG wild-type strain was utilized to reconstruct seven new mutants by single, double, and triple overexpression of the three enzymes key to this study. Subsequently, continuous cultures were performed to obtain steady-state metabolism in the organisms and experimental data were analyzed by metabolic flux analysis (MFA) to determine the regulation mechanisms. The MFA results showed that the seven overexpressed mutants all exhibited enhanced 2,3-BDO production, and the strain overexpressing the budBA gene produced the highest yield. While the enzyme encoded by the budA gene produced branched-chain amino acids which were favorable for cell growth, the budB gene enzyme rapidly enhanced the conversion of acetolactate to acetoin in an oxygen-dependent manner, and the budC gene enzyme catalyzed the reversible conversion of acetoin to 2,3-BDO and regulated the intracellular NAD(+)/NADH balance.

Observation of 2,3-butanediol biosynthesis in Lys regulator mutated Klebsiella pneumoniae at gene transcription level.

Microorganisms that produce 2,3-butanediol (2,3-BDO) are mostly mixed acid fermentation microorganisms, and they synthesize 2,3-BDO in order to suppress medium acidification. The 2,3-BDO operon (budBAC) is activated by the LysR regulator protein derived from the budR. In this study, the effect of the budR on 2,3-BDO-biosynthesis was observed at gene transcription level. The Klebsiella pneumoniae strains (wabG-deleted strain (SGSB100), budR over-expressed strain (SGSB101), and the budR-deleted strain (SGSB102)) were constructed. The resulting strains were cultivated in unified conditions. Samples were obtained at the lag-, log-, and stationary-phase of cell growth, and gene transcription levels of the budR, 2,3-BDO-biosynthesis-related (budB, budA, and budC), and acid-biosynthesis-related (ldhA and ack) genes were observed. During the lag-phase of cell growth in SGSB101, the budR transcription level increased approximately 8-fold, and 2,3-BDO production increased approximately 2-fold, when compared to SGSB100. Also in SGSB101 the transcription level of the acid-biosynthesis-related genes (ldhA and ack) increased approximately up to 11-fold during the lag-phase of cell growth compared to SGSB100. On contrast, in SGSB102 budR transcription was not detected, and the transcription level of the acid-biosynthesis-related genes (ldhA and ack) decreased approximately 70-fold during the lag-phase of cell growth compared to SGSB100. This is by far the first observation of 2,3-BDO regulation mechanism at gene transcription level in K. pneumoniae, and therefore may be useful for understanding and improving 2,3-BDO biosynthesis metabolic network.

Removal of pathogenic factors from 2,3-butanediol-producing Klebsiella species by inactivating virulence-related wabG gene.

Klebsiella species are the most extensively studied among a number of 2,3-butanediol (2,3-BDO)-producing microorganisms. The ability to metabolize a wide variety of substrates together with the ease of cultivation made this microorganisms particularly promising for the application in industrial-scale production of 2,3-BDO. However, the pathogenic characteristics of encapsulated Klebsiella species are considered to be an obstacle hindering their industrial applications. Here, we removed the virulence factors from three 2,3-BDO-producing strains, Klebsiella pneumoniae KCTC 2242, Klebsiella oxytoca KCTC1686, and K. oxytoca ATCC 43863 through site-specific recombination technique. We generated deletion mutation in wabG gene encoding glucosyltransferase which plays a key role in the synthesis of outer core lipopolysaccharides (LPS) by attaching the first outer core residue D-GalAp to the O-3 position of the L,D-HeppII residue. The morphologies and adhesion properties against epithelial cells were investigated, and the results indicated that the wabG mutant strains were devoid of the outer core LPS and lost the ability to retain capsular structure. The time profile of growth and 2,3-BDO production from K. pneumoniae KCTC 2242 and K. pneumoniae KCTC 2242 ΔwabG were analyzed in batch culture with initial glucose concentration of 70 g/l. The growth was not affected by disrupting wabG gene, but the production of 2,3-BDO decreased from 31.27 to 22.44 g/l in mutant compared with that of parental strain. However, the productions of acetoin and lactate from wabG mutant strain were negligible, whereas that from parental strain reached to ~5 g/l.

Enhanced activity of meso-secondary alcohol dehydrogenase from Klebsiella species by codon optimization.

Meso-secondary alcohol dehydrogenases (meso-SADH) from Klebsiella oxytoca KCTC1686 and Klebsiella pneumoniae KCTC2242 were codon optimized and expressed in Escherichia coli W3110. The published gene data of K. pneumoniae NTUH-K2044 (NCBI accession number AP006725), K. pneumoniae 342 (NCBI accession number CP000964), and K. pneumoniae MGH 78578 (NCBI accession number CP000647), were compared with the meso-SADH sequences of each strain, respectively. Codon-optimized meso-SADH enzymes of K. oxytoca and K. pneumoniae showed approximately twofold to fivefold increased enzyme activities for acetoin reduction over native enzymes. The highest activities for each strain were obtained at 30-37 °C and pH 6-7 (yielding 203.1 U/mg of protein and 156.5 U/mg of protein, respectively). The increased enzyme activity of the codon-optimized enzymes indicated that these modified enzymes could convert acetoin into 2,3-butanediol with a high yield.

Enhanced 2,3-butanediol production in recombinant Klebsiella pneumoniae via overexpression of synthesis-related genes.

2,3-Butanediol (2,3-BD) is a major metabolite produced by Klebsiella pneumoniae KCTC2242, which is a important chemical with wide applications. Three genes important for 2,3-BD biosynthesis acetolactate decarboxylase (budA), acetolactate synthase (budB), and alcohol dehydrogenase (budC) were identified in K. pneumoniae genomic DNA. With the goal of enhancing 2,3-BD production, these genes were cloned into pUC18K expression vectors containing the lacZ promoter and the kanamycin resistance gene to generate plasmids pSB1-7. The plasmids were then introduced into K. pneumoniae using electroporation. All strains were incubated in flask experiments and 2,3-BD production was increased by 60% in recombinant bacteria harboring pSB04 (budA and budB genes), compared with the parental strain K. pneumoniae KCTC2242. The maximum 2,3-BD production level achieved through fedbatch fermentation with K. pneumoniae SGJSB04 was 101.53 g/l over 40 h with a productivity of 2.54 g/l.h. These results suggest that overexpression of 2,3-BD synthesisrelated genes can enhance 2,3-BD production in K. pneumoniae by fermentation.

Identification of factors regulating Escherichia coli 2,3-butanediol production by continuous culture and metabolic flux analysis.

2,3-Butanediol (2,3-BDO) is an organic compound with a wide range of industrial applications. Although Escherichia coli is often used for the production of organic compounds, the wild-type E. coli does not contain two essential genes in the 2,3-BDO biosynthesis pathway, and cannot ferment 2,3-BDO. Therefore, a 2,3-BDO biosynthesis mutant strain of Escherichia coli was constructed and cultured. To determine the optimum culture factors for 2,3-BDO production, experiments were conducted under different culture environments ranging from strongly acidic to neutral pH. The extracellular metabolite profiles were obtained using high-performance liquid chromatography (HPLC), and the intracellular metabolite profiles were analyzed by ultra-performance liquid chromatography and quadruple time-of-flight mass spectrometry (UPLC/ Q-TOF-MS). Metabolic flux analysis (MFA) was used to integrate these profiles. The metabolite profiles showed that 2,3-BDO production favors an acidic environment (pH 5), whereas cell mass favors a neutral environment. Furthermore, when the pH of the culture fell below 5, both the cell growth and 2,3-BDO production were inhibited.

Synthesis of pure meso-2,3-butanediol from crude glycerol using an engineered metabolic pathway in Escherichia coli.

meso-2,3-Butanediol (meso-2,3-BDO) is essential for the synthesis of various economically valuable biosynthetic products; however, the production of meso-2,3-BDO from expensive carbon sources is an obstacle for industrial applications. In this study, genes involved in the synthesis of 2,3-BDO in Klebsiella pneumoniae were identified and used to genetically modify Escherichia coli for meso-2,3-BDO production. Two 2,3-BDO biosynthesis genes-budA, encoding acetolactate, and meso-budC, encoding meso-SADH-from K. pneumoniae were cloned into the pUC18 plasmid and introduced into E. coli. In 2 l batch culture, the SGSB03 E. coli strain yielded meso-2,3-BDO at 0.31 g/g(glucose) (with a maximum of 15.7 g/l(culture) after 48 h) and 0.21 g/g(crude glycerol) (with a maximum of 6.9 g/l(culture) after 48 h). Batch cultures were grown under optimized conditions (aerobic, 6% carbon source, 37 °C, and initial pH 7). To find the optimal culture conditions for meso-2,3-BDO production, we evaluated the enzyme activity of meso-SADH and the whole cell conversion yield (meso-2,3-BDO/acetoin) of the E. coli SGSB02, which contains pSB02. meso-SADH showed high enzyme activity at 30-37 °C and pH 7 (30.5-41.5 U/mg of protein), and the conversion yield of SGSB02 E. coli was highest at 37-42 °C and a pH of 7 (0.25-0.28 g( meso-2,3-BDO)/g(acetoin)).