TY  - JOUR
AU  - Wang, Qinghui
AU  - Gu, Jinjie
AU  - Shu, Lin
AU  - Jiang, Weiyan
AU  - Mojović, Ljiljana
AU  - Knežević-Jugović, Zorica
AU  - Shi, Jiping
AU  - Baganz, Frank
AU  - Lye, Gary J.
AU  - Xiang, Wensheng
AU  - Hao, Jian
PY  - 2022
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/5109
AB  - Klebsiella pneumoniae is a 2,3-butanediol producing bacterium. Nevertheless, a design and construction of l-valine production strain was studied in this paper. The first step of 2,3-butanediol synthesis and branched-chain amino acid synthesis pathways share the same step of α-acetolactate synthesis from pyruvate. However, the two pathways are existing in parallel and do not interfere with each other in the wild-type strain. A knockout of budA blocked the 2,3-butanediol synthesis pathway and resulted in the l-valine production. The budA coded an α-acetolactate decarboxylase and catalyzed the acetoin formation from α-acetolactate. Furthermore, blocking the lactic acid synthesis by knocking out of ldhA, which is encoding a lactate dehydrogenase, improved the l-valine synthesis. 2-Ketoisovalerate is the precursor of l-valine, it is also an intermediate of the isobutanol synthesis pathway, while indole-3-pyruvate decarboxylase (ipdC) is responsible for isobutyraldehyde formation from 2-ketoisovalerate. Production of l-valine has been improved by knocking out of ipdC. On the other side, the ilvE, encoding a transaminase B, reversibly transfers one amino group from glutamate to α-ketoisovalerate. Overexpression of ilvE exhibited a distinct improvement of l-valine production. The brnQ encodes a branched-chain amino acid transporter, and l-valine production was further improved by disrupting brnQ. It is also revealed that weak acidic and aerobic conditions favor l-valine production. Based on these findings, l-valine production by metabolically engineered K. pneumonia was examined. In fed-batch fermentation, 22.4 g/L of l-valine was produced by the engineered K. pneumoniae ΔbudA-ΔldhA-ΔipdC-ΔbrnQ-ilvE after 55 h of cultivation, with a substrate conversion ratio of 0.27 mol/mol glucose.
PB  - Springer Science and Business Media B.V.
T2  - World Journal of Microbiology and Biotechnology
T1  - Blocking the 2,3-butanediol synthesis pathway of Klebsiella pneumoniae resulted in l-valine production
IS  - 5
SP  - 81
VL  - 38
DO  - 10.1007/s11274-022-03266-9
ER  - 

@article{
author = "Wang, Qinghui and Gu, Jinjie and Shu, Lin and Jiang, Weiyan and Mojović, Ljiljana and Knežević-Jugović, Zorica and Shi, Jiping and Baganz, Frank and Lye, Gary J. and Xiang, Wensheng and Hao, Jian",
year = "2022",
abstract = "Klebsiella pneumoniae is a 2,3-butanediol producing bacterium. Nevertheless, a design and construction of l-valine production strain was studied in this paper. The first step of 2,3-butanediol synthesis and branched-chain amino acid synthesis pathways share the same step of α-acetolactate synthesis from pyruvate. However, the two pathways are existing in parallel and do not interfere with each other in the wild-type strain. A knockout of budA blocked the 2,3-butanediol synthesis pathway and resulted in the l-valine production. The budA coded an α-acetolactate decarboxylase and catalyzed the acetoin formation from α-acetolactate. Furthermore, blocking the lactic acid synthesis by knocking out of ldhA, which is encoding a lactate dehydrogenase, improved the l-valine synthesis. 2-Ketoisovalerate is the precursor of l-valine, it is also an intermediate of the isobutanol synthesis pathway, while indole-3-pyruvate decarboxylase (ipdC) is responsible for isobutyraldehyde formation from 2-ketoisovalerate. Production of l-valine has been improved by knocking out of ipdC. On the other side, the ilvE, encoding a transaminase B, reversibly transfers one amino group from glutamate to α-ketoisovalerate. Overexpression of ilvE exhibited a distinct improvement of l-valine production. The brnQ encodes a branched-chain amino acid transporter, and l-valine production was further improved by disrupting brnQ. It is also revealed that weak acidic and aerobic conditions favor l-valine production. Based on these findings, l-valine production by metabolically engineered K. pneumonia was examined. In fed-batch fermentation, 22.4 g/L of l-valine was produced by the engineered K. pneumoniae ΔbudA-ΔldhA-ΔipdC-ΔbrnQ-ilvE after 55 h of cultivation, with a substrate conversion ratio of 0.27 mol/mol glucose.",
publisher = "Springer Science and Business Media B.V.",
journal = "World Journal of Microbiology and Biotechnology",
title = "Blocking the 2,3-butanediol synthesis pathway of Klebsiella pneumoniae resulted in l-valine production",
number = "5",
pages = "81",
volume = "38",
doi = "10.1007/s11274-022-03266-9"
}

Wang, Q., Gu, J., Shu, L., Jiang, W., Mojović, L., Knežević-Jugović, Z., Shi, J., Baganz, F., Lye, G. J., Xiang, W.,& Hao, J.. (2022). Blocking the 2,3-butanediol synthesis pathway of Klebsiella pneumoniae resulted in l-valine production. in World Journal of Microbiology and Biotechnology
Springer Science and Business Media B.V.., 38(5), 81.
https://doi.org/10.1007/s11274-022-03266-9

Wang Q, Gu J, Shu L, Jiang W, Mojović L, Knežević-Jugović Z, Shi J, Baganz F, Lye GJ, Xiang W, Hao J. Blocking the 2,3-butanediol synthesis pathway of Klebsiella pneumoniae resulted in l-valine production. in World Journal of Microbiology and Biotechnology. 2022;38(5):81.
doi:10.1007/s11274-022-03266-9 .

Wang, Qinghui, Gu, Jinjie, Shu, Lin, Jiang, Weiyan, Mojović, Ljiljana, Knežević-Jugović, Zorica, Shi, Jiping, Baganz, Frank, Lye, Gary J., Xiang, Wensheng, Hao, Jian, "Blocking the 2,3-butanediol synthesis pathway of Klebsiella pneumoniae resulted in l-valine production" in World Journal of Microbiology and Biotechnology, 38, no. 5 (2022):81,
https://doi.org/10.1007/s11274-022-03266-9 . .

TY  - JOUR
AU  - Lu, Xiyang
AU  - Yao, Yao
AU  - Yang, Yang
AU  - Zhang, Zhongxi
AU  - Gu, Jinjie
AU  - Mojović, Ljiljana
AU  - Knežević-Jugović, Zorica
AU  - Baganz, Frank
AU  - Lye, Gary
AU  - Shi, Jiping
AU  - Hao, Jian
PY  - 2021
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/14
AB  - Ethylene glycol and glycolic acid are bulk chemicals with a broad range of applications. The ethylene glycol and glycolic acid biosynthesis pathways have been produced by microorganisms and used as a biological route for their production. Unlike the methods that use xylose or glucose as carbon sources, xylonic acid was used as a carbon source to produce ethylene glycol and glycolic acid in this study. Amounts of 4.2 g/L of ethylene glycol and 0.7 g/L of glycolic acid were produced by a wild-typeEscherichia coliW3110 within 10 H of cultivation with a substrate conversion ratio of 0.5 mol/mol. Furthermore,E. colistrains that produce solely ethylene glycol or glycolic acid were constructed. 10.3 g/L of glycolic acid was produced byE. coli Delta yqhD+aldA, and the achieved conversion ratio was 0.56 mol/mol. Similarly, theE. coli Delta aldA+yqhDproduced 8.0 g/L of ethylene glycol with a conversion ratio of 0.71 mol/mol. Ethylene glycol and glycolic acid production byE. colion xylonic acid as a carbon source provides new information on the biosynthesis pathway of these products and opens a novel way of biomass utilization.
PB  - Wiley, Hoboken
T2  - Biotechnology and Applied Biochemistry
T1  - Ethylene glycol and glycolic acid production by wild-typeEscherichia coli
EP  - 755
IS  - 4
SP  - 744
SP  - 755
VL  - 68
DO  - 10.1002/bab.1987
ER  - 

@article{
author = "Lu, Xiyang and Yao, Yao and Yang, Yang and Zhang, Zhongxi and Gu, Jinjie and Mojović, Ljiljana and Knežević-Jugović, Zorica and Baganz, Frank and Lye, Gary and Shi, Jiping and Hao, Jian",
year = "2021",
abstract = "Ethylene glycol and glycolic acid are bulk chemicals with a broad range of applications. The ethylene glycol and glycolic acid biosynthesis pathways have been produced by microorganisms and used as a biological route for their production. Unlike the methods that use xylose or glucose as carbon sources, xylonic acid was used as a carbon source to produce ethylene glycol and glycolic acid in this study. Amounts of 4.2 g/L of ethylene glycol and 0.7 g/L of glycolic acid were produced by a wild-typeEscherichia coliW3110 within 10 H of cultivation with a substrate conversion ratio of 0.5 mol/mol. Furthermore,E. colistrains that produce solely ethylene glycol or glycolic acid were constructed. 10.3 g/L of glycolic acid was produced byE. coli Delta yqhD+aldA, and the achieved conversion ratio was 0.56 mol/mol. Similarly, theE. coli Delta aldA+yqhDproduced 8.0 g/L of ethylene glycol with a conversion ratio of 0.71 mol/mol. Ethylene glycol and glycolic acid production byE. colion xylonic acid as a carbon source provides new information on the biosynthesis pathway of these products and opens a novel way of biomass utilization.",
publisher = "Wiley, Hoboken",
journal = "Biotechnology and Applied Biochemistry",
title = "Ethylene glycol and glycolic acid production by wild-typeEscherichia coli",
pages = "755-744-755",
number = "4",
volume = "68",
doi = "10.1002/bab.1987"
}

Lu, X., Yao, Y., Yang, Y., Zhang, Z., Gu, J., Mojović, L., Knežević-Jugović, Z., Baganz, F., Lye, G., Shi, J.,& Hao, J.. (2021). Ethylene glycol and glycolic acid production by wild-typeEscherichia coli. in Biotechnology and Applied Biochemistry
Wiley, Hoboken., 68(4), 744-755.
https://doi.org/10.1002/bab.1987

Lu X, Yao Y, Yang Y, Zhang Z, Gu J, Mojović L, Knežević-Jugović Z, Baganz F, Lye G, Shi J, Hao J. Ethylene glycol and glycolic acid production by wild-typeEscherichia coli. in Biotechnology and Applied Biochemistry. 2021;68(4):744-755.
doi:10.1002/bab.1987 .

Lu, Xiyang, Yao, Yao, Yang, Yang, Zhang, Zhongxi, Gu, Jinjie, Mojović, Ljiljana, Knežević-Jugović, Zorica, Baganz, Frank, Lye, Gary, Shi, Jiping, Hao, Jian, "Ethylene glycol and glycolic acid production by wild-typeEscherichia coli" in Biotechnology and Applied Biochemistry, 68, no. 4 (2021):744-755,
https://doi.org/10.1002/bab.1987 . .

TY  - JOUR
AU  - Yang, Chengyu
AU  - Fan, Weiming
AU  - Zhang, Ruijie
AU  - Shi, Jiping
AU  - Knežević-Jugović, Zorica
AU  - Zhang, Baoguo
PY  - 2019
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/4183
AB  - alpha-Arbutin (4-hydroquinone-alpha-D-glucopyranoside), an effective skin-lightening agent due to its considerable inhibitory effect on human tyrosinase activity, is widely used in the pharmaceutical and cosmetic industries. Recently, alpha-arbutin was prepared through transglucosylation of hydroquinone using microbial glycosyltransferases as catalysts. However, the low yield and prolonged reaction time of the biotransformation process of alpha-arbutin production limited its industrial application. In this work, an amylosucrase (ASase) from Xanthomonas campestris pv. campestris str. ATCC 33913 (XcAS) was expressed efficiently in Escherichia coli JM109. The catalytic property of the purified XcAS for the synthesis of alpha-arbutin was tested. The recombinant strain was applied for highly efficient synthesis of a-arbutin using sucrose and hydroquinone as glucosyl donor and acceptor, respectively. By optimizing the biotransformation conditions and applying a fed-batch strategy, the final production yield and conversion rate of alpha-arbutin reached 60.9 g/L and 95.5%, respectively, which is the highest reported yield by engineered strains. Compared to the highest reported value ( lt 1.4 g/L/h), our productivity (7.6 g/L/h) was improved more than five-fold. This work represents an efficient and rapid method for alpha-arbutin production with potential industrial applications.
PB  - MDPI, Basel
T2  - Catalysts
T1  - Study on Transglucosylation Properties of Amylosucrase from Xanthomonas campestris pv. Campestris and Its Application in the Production of alpha-Arbutin
IS  - 1
VL  - 9
DO  - 10.3390/catal9010005
ER  - 

@article{
author = "Yang, Chengyu and Fan, Weiming and Zhang, Ruijie and Shi, Jiping and Knežević-Jugović, Zorica and Zhang, Baoguo",
year = "2019",
abstract = "alpha-Arbutin (4-hydroquinone-alpha-D-glucopyranoside), an effective skin-lightening agent due to its considerable inhibitory effect on human tyrosinase activity, is widely used in the pharmaceutical and cosmetic industries. Recently, alpha-arbutin was prepared through transglucosylation of hydroquinone using microbial glycosyltransferases as catalysts. However, the low yield and prolonged reaction time of the biotransformation process of alpha-arbutin production limited its industrial application. In this work, an amylosucrase (ASase) from Xanthomonas campestris pv. campestris str. ATCC 33913 (XcAS) was expressed efficiently in Escherichia coli JM109. The catalytic property of the purified XcAS for the synthesis of alpha-arbutin was tested. The recombinant strain was applied for highly efficient synthesis of a-arbutin using sucrose and hydroquinone as glucosyl donor and acceptor, respectively. By optimizing the biotransformation conditions and applying a fed-batch strategy, the final production yield and conversion rate of alpha-arbutin reached 60.9 g/L and 95.5%, respectively, which is the highest reported yield by engineered strains. Compared to the highest reported value ( lt 1.4 g/L/h), our productivity (7.6 g/L/h) was improved more than five-fold. This work represents an efficient and rapid method for alpha-arbutin production with potential industrial applications.",
publisher = "MDPI, Basel",
journal = "Catalysts",
title = "Study on Transglucosylation Properties of Amylosucrase from Xanthomonas campestris pv. Campestris and Its Application in the Production of alpha-Arbutin",
number = "1",
volume = "9",
doi = "10.3390/catal9010005"
}

Yang, C., Fan, W., Zhang, R., Shi, J., Knežević-Jugović, Z.,& Zhang, B.. (2019). Study on Transglucosylation Properties of Amylosucrase from Xanthomonas campestris pv. Campestris and Its Application in the Production of alpha-Arbutin. in Catalysts
MDPI, Basel., 9(1).
https://doi.org/10.3390/catal9010005

Yang C, Fan W, Zhang R, Shi J, Knežević-Jugović Z, Zhang B. Study on Transglucosylation Properties of Amylosucrase from Xanthomonas campestris pv. Campestris and Its Application in the Production of alpha-Arbutin. in Catalysts. 2019;9(1).
doi:10.3390/catal9010005 .

Yang, Chengyu, Fan, Weiming, Zhang, Ruijie, Shi, Jiping, Knežević-Jugović, Zorica, Zhang, Baoguo, "Study on Transglucosylation Properties of Amylosucrase from Xanthomonas campestris pv. Campestris and Its Application in the Production of alpha-Arbutin" in Catalysts, 9, no. 1 (2019),
https://doi.org/10.3390/catal9010005 . .

TY  - JOUR
AU  - Sun, Yuehong
AU  - Wei, Dong
AU  - Shi, Jiping
AU  - Mojović, Ljiljana
AU  - Han, Zengsheng
AU  - Hao, Jian
PY  - 2014
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/2832
AB  - 2-Ketogluconic acid production by Klebsiella pneumoniae is a pH-dependent process, strictly proceeding under acidic conditions. Unfortunately, cell growth is inhibited by acidic conditions, resulting in low productivity of 2-ketogluconic acid. To overcome this deficiency, a two-stage fermentation strategy was exploited in the current study. During the first stage, the culture was maintained at neutral pH, favoring cell growth. During the second stage, the culture pH was switched to acidic conditions favoring 2-ketogluconic acid accumulation. Culture parameters, including switching time, dissolved oxygen levels, pH, and temperature were optimized for the fed-batch fermentation. Characteristics of glucose dehydrogenase and gluconate dehydrogenase were revealed in vitro, and the optimal pHs of the two enzymes coincided with the optimum culture pH. Under optimum conditions, a total of 186 g/l 2-ketogluconic acid was produced at 26 h, and the conversion ratio was 0.98 mol/mol. This fermentation strategy has successfully overcome the mismatch between optimum parameters required for cell growth and 2-ketogluconic acid accumulation, and this result has the highest productivity and conversion ratio of 2-ketogluconic and produced by microorganism.
PB  - Korean Soc Microbiology & Biotechnology, Seoul
T2  - Journal of Microbiology and Biotechnology
T1  - Two-Stage Fermentation for 2-Ketogluconic Acid Production by Klebsiella pneumoniae
EP  - 787
IS  - 6
SP  - 781
VL  - 24
DO  - 10.4014/jmb.1401.01038
ER  - 

@article{
author = "Sun, Yuehong and Wei, Dong and Shi, Jiping and Mojović, Ljiljana and Han, Zengsheng and Hao, Jian",
year = "2014",
abstract = "2-Ketogluconic acid production by Klebsiella pneumoniae is a pH-dependent process, strictly proceeding under acidic conditions. Unfortunately, cell growth is inhibited by acidic conditions, resulting in low productivity of 2-ketogluconic acid. To overcome this deficiency, a two-stage fermentation strategy was exploited in the current study. During the first stage, the culture was maintained at neutral pH, favoring cell growth. During the second stage, the culture pH was switched to acidic conditions favoring 2-ketogluconic acid accumulation. Culture parameters, including switching time, dissolved oxygen levels, pH, and temperature were optimized for the fed-batch fermentation. Characteristics of glucose dehydrogenase and gluconate dehydrogenase were revealed in vitro, and the optimal pHs of the two enzymes coincided with the optimum culture pH. Under optimum conditions, a total of 186 g/l 2-ketogluconic acid was produced at 26 h, and the conversion ratio was 0.98 mol/mol. This fermentation strategy has successfully overcome the mismatch between optimum parameters required for cell growth and 2-ketogluconic acid accumulation, and this result has the highest productivity and conversion ratio of 2-ketogluconic and produced by microorganism.",
publisher = "Korean Soc Microbiology & Biotechnology, Seoul",
journal = "Journal of Microbiology and Biotechnology",
title = "Two-Stage Fermentation for 2-Ketogluconic Acid Production by Klebsiella pneumoniae",
pages = "787-781",
number = "6",
volume = "24",
doi = "10.4014/jmb.1401.01038"
}

Sun, Y., Wei, D., Shi, J., Mojović, L., Han, Z.,& Hao, J.. (2014). Two-Stage Fermentation for 2-Ketogluconic Acid Production by Klebsiella pneumoniae. in Journal of Microbiology and Biotechnology
Korean Soc Microbiology & Biotechnology, Seoul., 24(6), 781-787.
https://doi.org/10.4014/jmb.1401.01038

Sun Y, Wei D, Shi J, Mojović L, Han Z, Hao J. Two-Stage Fermentation for 2-Ketogluconic Acid Production by Klebsiella pneumoniae. in Journal of Microbiology and Biotechnology. 2014;24(6):781-787.
doi:10.4014/jmb.1401.01038 .

Sun, Yuehong, Wei, Dong, Shi, Jiping, Mojović, Ljiljana, Han, Zengsheng, Hao, Jian, "Two-Stage Fermentation for 2-Ketogluconic Acid Production by Klebsiella pneumoniae" in Journal of Microbiology and Biotechnology, 24, no. 6 (2014):781-787,
https://doi.org/10.4014/jmb.1401.01038 . .