Comparative study of immunobiological properties of Klebsiella planticola and Klebsiella pneumoniae strains

The present investigation was based on information on the frequent isolation of K. planticola, inhabiting soil and water ecosystems, from respiratory tract secretions, urine, wound surface and other clinical material in infectious pathology of humans. The comparative study of the main immunobiological properties of K. planticola strain TCXA 91 and K. pneumoniae strain 204 was carried out. The strains were equally virulent for white mice, the infective dose being about 100x10(6) when introduced intraperitoneally in 0.4% agar. These strains exhibited protective cross activity in the animals challenged with K. pneumoniae virulent strain K16. The survival rates of mice for K. planticola TCXA 91 and K. pneumoniae 204 were, respectively, 60% and 70%, the death rate of the control animals being 100%. For the first time K. pneumoniae 204 was found capable of stimulating the growth of lawn grasses in the presence of sodium chloride. The results were evaluated by the germination of seeds, the length of the plantlet and the root. The data obtained in this study are indicative of the similarity of the immunobiological properties of K. planticola TCXA 91 and K. pneumoniae 204.     

Lipopolysaccharide O1 antigen contributes to the virulence in Klebsiella pneumoniae causing pyogenic liver abscess

Klebsiella pneumoniae is the common cause of a global emerging infectious disease, community-acquired pyogenic liver abscess (PLA). Capsular polysaccharide (CPS) and lipopolysaccharide (LPS) are critical for this microorganism's ability to spread through the blood and to cause sepsis. While CPS type K1 is an important virulence factor in K. pneumoniae causing PLA, the role of LPS in PLA is not clear. Here, we characterize the role of LPS O antigen in the pathogenesis of K. pneumoniae causing PLA. NTUH-K2044 is a LPS O1 clinical strain; the presence of the O antigen was shown via the presence of 1,3-galactan in the LPS, and of sequences that align with the wb gene cluster, known to produce O-antigen. Serologic analysis of K. pneumoniae clinical isolates demonstrated that the O1 serotype was more prevalent in PLA strains than that in non-tissue-invasive strains (38/42 vs. 9/32, P<0.0001). O1 serotype isolates had a higher frequency of serum resistance, and mutation of the O1 antigen changed serum resistance in K. pneumoniae. A PLA-causing strain of CPS capsular type K2 and LPS serotype O1 (i.e., O1:K2 PLA strain) deleted for the O1 synthesizing genes was profoundly attenuated in virulence, as demonstrated in separate mouse models of septicemia and liver abscess. Immunization of mice with the K2044 magA-mutant (K(1) (-) O(1)) against LPS O1 provided protection against infection with an O1:K2 PLA strain, but not against infection with an O1:K1 PLA strain. Our findings indicate that the O1 antigen of PLA-associated K. pneumoniae contributes to virulence by conveying resistance to serum killing, promoting bacterial dissemination to and colonization of internal organs after the onset of bacteremia, and could be a useful vaccine candidate against infection by an O1:K2 PLA strain.     

2,3-丁二醇代谢途径关键酶基因敲除对克雷伯氏菌发酵产1,3-丙二醇的影响

2,3-丁二醇是克雷伯氏菌发酵产1,3-丙二醇的主要副产物,为减少2,3-丁二醇的产生,利用Red重组技术对克雷伯氏菌2,3-丁二醇合成途径关键酶基因budC和budA进行了敲除。突变株发酵性能实验结果表明,所获得的两株突变株生长性能受到不同程度的影响;budC基因的缺失使菌株1,3-丙二醇产量提高了10%,2,3-丁二醇降低为原来的70%,而budA基因缺失则使菌株无2,3-丁二醇和1,3-丙二醇的产生,但乳酸、琥珀酸、乙醇和乙酸的产量较出发菌株都有明显增长。通过进一步对budC基因缺失菌株主要产物分析,推测在该菌中存在2,3-丁二醇回补途径,这一结果为低副产物克雷伯氏菌的改造提供了新依据。     

Effect of metal oxides on the growth,hemolytic and serologic properties of Klebsiella pneumoniae

Silicon, dysprosium, germanium, yttrium, iron, cobalt, samarium, lutecium oxides, as well as the mixture of 8 metal oxides, at a concentration of 20 g/l were found to produce a stimulating or inhibiting effect on the growth of K. pneumoniae strains 204 and K-9. Silicon, dysprosium, germanium and yttrium oxides were shown to stimulate the growth of K. pneumoniae strain 204. Iron, cobalt, samarium and lutecium oxides, as well as the mixtures of all oxides under study, inhibited the growth of this strain. Silicon, samarium and lutecium oxides produced no effect on the growth of K. pneumoniae strain K-9; at the same time germanium and yttrium oxides stimulated the growth of these bacteria, while dysprosium, iron, cobalt oxides, as well as the mixture of all oxides, inhibited their growth. The presence of metal oxides did not change the serological activity of the cultures of both strains growing old, i.e. by 24 hours of their growth. The addition of silicon, germanium and iron oxides to the culture medium increased the hemolytic activity of K. pneumoniae strain K-9 seven to ninefold in comparison with the control grown in a synthetic nutrient medium without metal oxides. The comparison of these two strains (K-9 and 204) revealed that K. pneumoniae strain K-9 possessed greater hemolytic activity.     

Behavior of a hybrid F' ts114 lac+, his+ factor (F42-400) in Klebsiella pneumoniae M5a1.

Episome F' ts114 lac+, his+ (F42-400) was transferred from Salmonella typhimurium to Klebsiella pneumoniae. From the progeny, a strain of K. pneumoniae able to retransfer the episome was obtained. The His+ phenotype in this strain is temperature sensitive. Escherichia coli female-specific phages phiII, W31, and T3 were shown to plate on K. pneumoniae. From phiII we obtained two derivatives; phiIIK, which plates only on K. pneumoniae, and phiIIE, which plates only on E. coli. Growth of phages T3 and phiIIK was inhibited by F42-400 in K. pneumoniae. Growth in presence of acridine orange in a defined medium at 40 C resulted in a high level of curing. The frequency of His+ cells after growth in acridine orange at 40 C was 0.001%. An extensive search to detect chromosome mobilization by F42-400 in K. pneumoniae, under different experimental conditions, was negative. We cannot exclude the possibility that the low transfer efficiencies prevented our detection of chromosome mobilization. A search among temperature-resistant, acridine orange-curing-resistant, or galactose-resistant derivatives of the K. pneumoniae donor strain failed to reveal any chromosome transfer. Our failure to detect Hfr's may be a result of: (i) the peculiarity of episome F42-400, (ii) the peculiarity of K. pneumoniae chromosome, or (iii) low transfer efficiency. K. pneumoniae-modified F42-400 and phage 424 were restricted by E. Coli K-12. E. coli K-12-modified episome F42-400 and phage 424 were restricted by K. pneumoniae. E. coli C failed to restrict F42-400 modified with K. pneumoniae specificity. The ability of K. pneumoniae to accept F42-400 is less, by about a factor of 50, than that of E. coli C. As an explanation for the differences in the behavior of E. coli C and K. pneumoniae in ability to receive F42-400 it was suggested that recipient bacteria have specific sites for interaction with the F-pilus tip; these are present in E. Coli C, leading to high transfer efficiency, whereas they may not be present (or if present, are not accessible) in K. pneumoniae, leading to low transfer efficiency.     

Preliminary investigation of a mice model of Klebsiella pneumoniae subsp. ozaenae induced pneumonia

In the present study, we comparatively assessed the pathophysiological mechanisms developed during lung infection of BALB/C female mice infected by an original wild type Klebsiella pneumoniae subsp. ozaenae strain (CH137) or by a referent subspecies K. pneumoniae. subsp. pneumoniae strain (ATCC10031). The mice infected with 2.10? CFU K. p. subsp. pneumoniae (n = 10) showed transient signs of infection and all of them recovered. All of those infected with 1.10? CFU K. p. subsp. ozaenae (n = 10) developed pneumonia within 24 h and died between 48 and 72 h. Few macrophages, numerous polymorphonuclear cells and lymphocytes were observed in their lungs in opposite to K. p. subsp. pneumoniae. In bronchoalveolar lavage, a significant increase in MIP-2, IL-6, KC and MCP-1 levels was only observed in K. p. subsp. ozaenae infected mice whereas high levels of TNF-α were evidenced with the two subspecies. Our findings indicated a lethal effect of a wild type K. p. subsp. ozaenae strain by acute pneumonia reflecting an insufficient alveolar macrophage response. This model might be of a major interest to comparatively explore the pathogenicity of K. p. subsp ozaenae strains and to further explore the physiopathological mechanisms of gram-negative bacteria induced human pneumonia.     

Novel aerobactin receptor in Klebsiella pneumoniae

Several Klebsiella pneumoniae strains which produced enterochelin but not aerobactin were nevertheless sensitive to cloacin DF13. In contrast, a strain of serotype K1:O1 which produced both siderophores was cloacin-resistant. Loss by mutation of the O1 but not K1 antigen rendered this strain cloacin-sensitive, indicating that the O1 antigen prevented access of cloacin to the cloacin/aerobactin receptor. Unlike the K1:O1 strain, the aerobactin-negative strains failed to hybridize in a colony blot assay with an aerobactin receptor gene probe prepared from pColV-K30. However, antisera raised against the 74 kDa pColV-K30 aerobactin receptor cross-reacted with a 76 kDa outer-membrane protein in each K. pneumoniae strain. In addition to the 76 kDa protein, the K1:O1 strain also produced a strongly cross-reacting 74 kDa protein. To determine whether these aerobactin-negative strains could use aerobactin, mutants unable to synthesize siderophores were isolated. Aerobactin promoted the growth of these mutants in iron-deficient media. The evidence presented suggests that some K. pneumoniae strains produce an aerobactin iron-uptake system without apparent production of aerobactin and which is probably based on a 76 kDa receptor, the gene for which does not hybridize with aerobactin receptor gene encoded on pColV-K30.     

聚羟基丁酸路径在克雷伯氏菌中的构建

以生物柴油的副产物甘油生产高附加值的1,3-丙二醇,现已成为提升生物柴油产业链经济性的重要途径,而中间代谢产物3-羟基丙醛积累造成细胞死亡,发酵异常终止是生物法生产1,3-丙二醇过程中的关键问题。不同于传统的降低3-羟基丙醛积累的思路,本文从增强克雷伯氏菌对3-羟基丙醛的抗逆性出发,改善克雷伯氏菌1,3-丙二醇的生产性能,首次将聚羟基丁酸路径引入克雷伯氏菌中,构建了新型基因工程菌,并对其1,3-丙二醇发酵性能及聚羟基丁酸代谢进行了初步的研究。经IPTG诱导,工程菌中检测到聚羟基丁酸,其含量随IPTG浓度增加而增大。优化的IPTG浓度为0.5 mmol/L。初始甘油50 g/L时,野生菌可正常发酵生产1,3-丙二醇,1,3-丙二醇浓度达到22.1 g/L,其质量得率为46.4%。当初始甘油达到70 g/L时,由于高浓度3-HPA积累,野生菌发酵终止,而工程菌可正常发酵生产1,3-丙二醇,PDO产量可达31.3 g/L,其质量得率为43.9%。同时检测到聚羟基丁酸积累。研究结果有助于加深对克雷伯氏菌1,3-丙二醇代谢机理的认识,为克雷伯氏菌的进一步优化提供了新的思路。     

Regulation of Nitrogen Fixation in Klebsiella pneumoniae: Evidence for a Role of Glutamine Synthetase as a Regulator of Nitrogenase Synthesis

Mutations causing constitutive synthesis of glutamine synthetase (GlnC(-) phenotype) were transferred from Klebsiella aerogenes into Klebsiella pneumoniae by P1-mediated transduction. Such GlnC(-) strains of K. pneumoniae have constitutive levels of glutamine synthetase. Two of three GlnC(-) strains of K. pneumoniae studied, each containing independently isolated mutations that confer the GlnC(-) phenotype, continue to synthesize nitrogenase in the presence of NH(4) (+). One strain, KP5069, produces 30% as much nitrogenase when grown in the presence of 15 mM NH(4) (+) as in its absence. The GlnC(-) phenotype allows the synthesis of nitrogenase to continue under conditions that completely repress nitrogenase synthesis in the wild-type strain. Glutamine auxotrophs of K. pneumoniae, that do not produce catalytically active glutamine synthetase, are unable to synthesize nitrogenase during nitrogen limited growth. Complementation of K. pneumoniae Gln(-) strains by an Escherichia coli episome (F'133) simultaneously restores glutamine synthetase activity and the ability to synthesize nitrogenase. These results indicate a role for glutamine synthetase as a positive control element for nitrogen fixation in K. pneumoniae.     

利用甘油发酵耦联生产3-羟基丙酸及1,3-丙二醇重组菌的构建及筛选

在肺炎克雷伯杆菌(Klebsiella pneumoniae)代谢甘油生产1,3-丙二醇(1,3-PD)的过程中,为了减少有毒中间产物3-羟基丙醛(3-HPA)的积累,可将其转化为3-羟基丙酸(3-HP),从而实现1,3-丙二醇和3-羟基丙酸的联产。克隆来自于酿酒酵母的NAD+依赖型的乙醛脱氢酶(ALDH)的基因aldh4,构建了表达载体pKP-aldh,转化K.pneumoniae,得到了有效表达乙醛脱氢酶的重组肺炎克雷伯杆菌(K.pneumoniae A+)。在此基础上,使用紫外诱变联合菌种驯化的方法对K.pneumoniae A+进行筛选,获得了可耐受较高3-HP浓度(≥35 g/L)的重组肺炎克雷伯杆菌K.pneumoniae A+5-3。发酵实验结果表明,K.pneumoniae A+5-3可将3-HPA转化为3-HP,能够同时利用甘油耦联生产3-HP和1,3-PD,产量分别达到5.0 g/L和74.5 g/L。     

Application of PCR ribotyping and tDNA-PCR for Klebsiella pneumoniae identification

PCR analysis of 16S-23S internal transcribed spacer (PCR ribotyping) and tRNA intergenic spacer (tDNA-PCR) were evaluated for their effectiveness in identification of clinical strains of Klebsiella pneumoniae and differentiation with related species. For this purpose both methods were applied to forty-three clinical isolates biochemically identified as K. pneumoniae subsp. pneumoniae isolated from patients clinical specimens attended at five hospitals in three Brazilian cities. References strains of K. pneumoniae subsp. pneumoniae, K. pneumoniae subsp. ozaenae, K. oxytoca, K. planticola and Enterobacter aerogenes were also analyzed. Both PCR methods showed specific patterns for each species. A conserved PCR ribotype pattern was observed for all clinical K. pneumoniae isolates, while differing from other related analyzed species. tDNA-PCR revealed five distinct patterns among the K. pneumoniae clinical isolates studied, demonstrating a predominant group with 90.6% of isolates presenting the same pattern of K. pneumoniae type strain. Both PCR-based methods were not able to differentiate K. pneumoniae subspecies. On the basis of the results obtained, both methods were efficient to differentiate the Klebsiella species analyzed, as well as E. aerogenes. Meanwhile tDNA-PCR revealed different tRNA arrangements in K. pneumoniae, suggesting intra-species heterogeneity of their genome organization, the polymorphism of the intergenic spacers between 16S and 23S rRNA genes appears to be highly conserved whithin K. pneumoniae clinical isolates, showing that PCR ribotyping can be an useful tool for identification of K. pneumoniae isolates.     

1997-2010年小儿肺炎克雷伯菌耐药性变迁分析

目的 了解1997-2010年肺炎克雷伯菌(KPN)在儿科及新生儿科临床感染中的流行状况及耐药性.方法 收集珠海市妇幼保健院儿科及新生儿科1997-2010年临床痰液标本中分离的肺炎克雷伯菌共633株,分析其对常用抗生素的耐药性.并分析2005年以后产超广谱β-内酰胺酶(ESBLs)与不产ESBLs肺炎克雷伯菌株耐药性的差异.结果 根据14年来该院临床分离的肺炎克雷伯菌药敏试验结果显示,该菌对氨苄西林及哌拉西林的耐药率最高,可达100％.对头孢他啶、头孢吡肟的耐药率各年有波动,但总体无明显变化(分别约28％、34％).对儿科临床限制使用药物如庆大霉素、左旋氧氟沙星、氯霉素、复方新诺明等耐药率呈逐年下降趋势.14年来尚未发现对亚胺培南耐药的KPN.2005年以后产ESBLs菌株的检出率呈逐年增高趋势(17％ ～38.7％),且对各种抗生素的耐药率较普通KPN明显增高.结论 肺炎克雷伯菌对临床常用β-内酰胺类抗生素耐药率高,儿科限制使用药物的耐药率有逐年下降趋势,产ESBLS菌株耐药情况严重.     

Contribution of capsular and lipopolysaccharide antigens to the pathogenesis of<Emphasis Type="Italic">Klebsiella pneumoniae</Emphasis> respiratory tract infection

The role of Klebsiella pneumoniae K- and O-polysaccharide antigens was determined in a rat model of lobar pneumonia. The induction of experimental infection in rats by wild-type strain, and its lipopolysaccharide- and capsular polysaccharide-deficient mutants was compared. Though the mutant lacking both antigens (K- O-) induced infection it could not successfully establish itself in the rat lung. It caused only mild infection, as compared to the wild type strain (K+ O+) and the strain lacking CPS alone (K- O+). Besides capsular polysaccharide, the lipopolysaccharide antigen was shown to be an important factor in pathogenesis of K. pneumoniae acute respiratory tract infection.     

Identification of carbapenems resistant genes on biofilm forming K. pneumoniae from urinary tract infection

The multi-drug resistant effect of the Gram negative bacteria K. pneumoniae was identified by disc diffusion method using specific UTI panel discs of Kleb 1 HX077 and Kleb 2 HX090 HEXA. Among the multi-drug resistant bacteria, the carbapenem resistant (CR) effect of the K. pneumoniae was screened by specific carbapenem detection antibiotics of HEXA HX066 and HX0103 HEXA by disc diffusion method. In addition, the effective antibiotics were further performed against K. pneumoniae by minimum inhibition concentration method. Further, the carbapenemase genes of VIM 1 and IMP 1 were detected from the isolated strains by multiplex PCR method. Furthermore, the biofilm forming ability of selected carbapenem resistant K. pneumoniae was initially identified by tissue culture plate method and confirmed by exopolysaccharide arrest ability of congo red agar assay. Finally, our result was proved that the identified K. pneumoniae is carbapenemase producing strain, and its virulence was extended with strong biofilm formation.     

Histamine production by Klebsiella pneumoniae and an incident of scombroid fish poisoning.

A histamine-producing strain of Klebsiella pneumoniae was isolated from a sample of tuna sashimi implicated in an outbreak of scombroid fish poisoning. None of the other nine gram-negative bacterial strains isolated from the tuna sashimi was capable of equivalent histamine production. Bacterial histamine production was monitored in a tuna fish infusion broth (TFIB), and the implicated K. pneumoniae was capable of producing 442 mg of histamine per 100 g of tuna in TFIB in 7 h under controlled incubation conditions. Only 12 of 50 other K. pneumoniae strains, representing 5 distinct biochemical types, which had been originally isolated from foods, were able to produce such levels of histamine in TFIB. No correlation was found between histamine production and other biochemical characteristics or antibiotic resistance. Of the 12 histamine-producing strains, 11 belonged to type 2, which is characterized as indole negative with positive reactions in the urea and Voges-Proskauer tests. However, only 50% of the type 2 strains examined produced high levels of histamine in TFIB. Additionally, the implicated K. pneumoniae strain and one other strain belonged to type 1, which is characterized by positive reactions in the indole, urea, and Voges-Proskauer tests.     

Histamine production by Klebsiella pneumoniae and an incident of scombroid fish poisoning.

A histamine-producing strain of Klebsiella pneumoniae was isolated from a sample of tuna sashimi implicated in an outbreak of scombroid fish poisoning. None of the other nine gram-negative bacterial strains isolated from the tuna sashimi was capable of equivalent histamine production. Bacterial histamine production was monitored in a tuna fish infusion broth (TFIB), and the implicated K. pneumoniae was capable of producing 442 mg of histamine per 100 g of tuna in TFIB in 7 h under controlled incubation conditions. Only 12 of 50 other K. pneumoniae strains, representing 5 distinct biochemical types, which had been originally isolated from foods, were able to produce such levels of histamine in TFIB. No correlation was found between histamine production and other biochemical characteristics or antibiotic resistance. Of the 12 histamine-producing strains, 11 belonged to type 2, which is characterized as indole negative with positive reactions in the urea and Voges-Proskauer tests. However, only 50% of the type 2 strains examined produced high levels of histamine in TFIB. Additionally, the implicated K. pneumoniae strain and one other strain belonged to type 1, which is characterized by positive reactions in the indole, urea, and Voges-Proskauer tests.     

Protective activity of a cell-free Klebsiella vaccine in relation to different Klebsiella pneumoniae serovars

The capacity of dried Klebsiella cell-free vaccine, obtained from strain No. 204 by the disintegration of microbial cells with hydroxylamine, for protecting mice from Klebsiella septic infection caused by the homologous serovar and 9 heterologous serovars of K. pneumoniae was studied. The newly developed preparation was found capable of stimulating immunity not only to the homologous K. pneumoniae serovar, but also to other K. pneumoniae heterologous serovars: K1, K9, K11, K16, K20, K61. The protective capacity of the preparation with respect to these serovars was not inferior to that of the vaccines prepared by the same method from the corresponding homologous strains. The capacity of the vaccine to protect mice from Klebsiella sepsis was manifested irrespective of the virulence of the strains used for challenge.     

Protease activity of Klebsiella pneumoniae of different virulence

The study of substrate specificity and activity of proteolytic enzymes secreted by K. pneumoniae strains with different virulence was carried out. The strains were cultivated in a liquid semi-synthetic medium. The biomass was inactivated, and the supernatant fluid was separated from microbial cells by centrifuging. In the supernatant thus obtained and in the fractions isolated by gel filtration with the subsequent purification on DEAE Sepharose elastase-like, trypsin-like and chemotrypsin-like proteolytic activity was determined. In K. pneumoniae strains with different virulence only a single proteolytic enzyme--elastase with a mol. wt. of 21 kD--was detected. The protease activity of the supernatant culture fluid did not depend on the virulence of the strain and was equal to 5,416-7,476 I.U./ml. The activity of the purified enzyme was 100% of the elastase-like activity of the supernatant culture fluid. The most virulent K. pneumoniae strain K2, whose LD50 for white mice was less than 10 microbial cells, was characterized by lower elastase-like activity. The absence of correlation between protease activity and K. pneumoniae virulence may be explained by the fact that surface glycoproteins of eukaryotic cells are glycosilated and thus slightly accessible for proteases.     

Host and microbiota factors that control Klebsiella pneumoniae mucosal colonization in mice

Klebsiella pneumoniae is both an opportunistic pathogen and a commensal organism. We have previously reported that K. pneumoniae strain IA565 (KpIA565) is non-pathogenic in a murine model of acute pneumonia. In this study, KpIA565 was inoculated into wild-type mice and found to stably colonize and persist in the nasal cavity and gastrointestinal tract of mice for up to 3weeks post-inoculation. Intranasal inoculation of wild-type or germ-free mice with KpIA565 resulted in similar bacterial levels in the nasal cavity, suggesting KpIA565 nasal colonization is independent of normal nasal microbiota. In contrast, KpIA565 gastrointestinal tract colonization was significantly higher in germ-free mice than in wild-type mice, indicating that members of the endogenous microbiota regulate KpIA565 colonization. In the presence of non-specific dextran sodium sulfate-induced inflammation, KpIA565 gastrointestinal tract colonization was significantly higher when compared to non-DSS treated mice. Interestingly, KpIA565 colonization was unaffected by Citrobacter rodentium-induced gastrointestinal tract inflammation. However, gastrointestinal tract colonization with K. pneumoniae strain IA565 had no impact on the inflammatory histopathology in either colitis model. This study is the first to identify and describe mechanisms influencing the growth and behavior of a murine commensal strain of K. pneumoniae.     

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.