Production of α-ketoisovalerate with whey powder by systemic metabolic engineering of Klebsiella oxytoca.
Klebsiella oxytoca
L-Valine
Metabolic engineering
Whey powder
α-Ketoisovalerate
Journal
Microbial cell factories
ISSN: 1475-2859
Titre abrégé: Microb Cell Fact
Pays: England
ID NLM: 101139812
Informations de publication
Date de publication:
05 Oct 2024
05 Oct 2024
Historique:
received:
20
07
2024
accepted:
29
09
2024
medline:
5
10
2024
pubmed:
5
10
2024
entrez:
4
10
2024
Statut:
epublish
Résumé
Whey, which has high biochemical oxygen demand and chemical oxygen demand, is mass-produced as a major by-product of the dairying industry. Microbial fermentation using whey as the carbon source may convert this potential pollutant into value-added products. This study investigated the potential of using whey powder to produce α-ketoisovalerate, an important platform chemical. Klebsiella oxytoca VKO-9, an efficient L-valine producing strain belonging to Risk Group 1 organism, was selected for the production of α-ketoisovalerate. The leucine dehydrogenase and branched-chain α-keto acid dehydrogenase, which catalyzed the reductive amination and oxidative decarboxylation of α-ketoisovalerate, respectively, were inactivated to enhance the accumulation of α-ketoisovalerate. The production of α-ketoisovalerate was also improved through overexpressing α-acetolactate synthase responsible for pyruvate polymerization and mutant acetohydroxyacid isomeroreductase related to α-acetolactate reduction. The obtained strain K. oxytoca KIV-7 produced 37.3 g/L of α-ketoisovalerate from lactose, the major utilizable carbohydrate in whey. In addition, K. oxytoca KIV-7 also produced α-ketoisovalerate from whey powder with a concentration of 40.7 g/L and a yield of 0.418 g/g. The process introduced in this study enabled efficient α-ketoisovalerate production from low-cost substrate whey powder. Since the key genes for α-ketoisovalerate generation were integrated in genome of K. oxytoca KIV-7 and constitutively expressed, this strain is promising in stable α-ketoisovalerate fermentation and can be used as a chassis strain for α-ketoisovalerate derivatives production.
Sections du résumé
BACKGROUND
BACKGROUND
Whey, which has high biochemical oxygen demand and chemical oxygen demand, is mass-produced as a major by-product of the dairying industry. Microbial fermentation using whey as the carbon source may convert this potential pollutant into value-added products. This study investigated the potential of using whey powder to produce α-ketoisovalerate, an important platform chemical.
RESULTS
RESULTS
Klebsiella oxytoca VKO-9, an efficient L-valine producing strain belonging to Risk Group 1 organism, was selected for the production of α-ketoisovalerate. The leucine dehydrogenase and branched-chain α-keto acid dehydrogenase, which catalyzed the reductive amination and oxidative decarboxylation of α-ketoisovalerate, respectively, were inactivated to enhance the accumulation of α-ketoisovalerate. The production of α-ketoisovalerate was also improved through overexpressing α-acetolactate synthase responsible for pyruvate polymerization and mutant acetohydroxyacid isomeroreductase related to α-acetolactate reduction. The obtained strain K. oxytoca KIV-7 produced 37.3 g/L of α-ketoisovalerate from lactose, the major utilizable carbohydrate in whey. In addition, K. oxytoca KIV-7 also produced α-ketoisovalerate from whey powder with a concentration of 40.7 g/L and a yield of 0.418 g/g.
CONCLUSION
CONCLUSIONS
The process introduced in this study enabled efficient α-ketoisovalerate production from low-cost substrate whey powder. Since the key genes for α-ketoisovalerate generation were integrated in genome of K. oxytoca KIV-7 and constitutively expressed, this strain is promising in stable α-ketoisovalerate fermentation and can be used as a chassis strain for α-ketoisovalerate derivatives production.
Identifiants
pubmed: 39367476
doi: 10.1186/s12934-024-02545-4
pii: 10.1186/s12934-024-02545-4
doi:
Substances chimiques
alpha-ketoisovalerate
759-05-7
Hemiterpenes
0
Powders
0
Acetolactate Synthase
EC 2.2.1.6
Keto Acids
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
264Subventions
Organisme : National Natural Science Foundation of China
ID : 32300045
Organisme : the National Key R&D Program of China
ID : 2022YFC2106000
Organisme : Youth Program of Natural Science Foundation of Shandong Province
ID : ZR2022QC092
Organisme : China Postdoctoral Science Foundation
ID : 2023M742085
Organisme : Youth Program of Natural Science Foundation of Qingdao City
ID : 23-2-1-31-zyyd-jch
Organisme : Postdoctoral Innovation Program of Shandong Province
ID : No. SDCX-ZG-202400150
Organisme : Qingdao Postdoctoral Research Project
ID : QDBSH20230201011
Organisme : State Key Laboratory of Microbial Technology Open Projects Fund
ID : M2023-02
Organisme : State Key Laboratory of Microbial Technology
ID : SKLMTFCP-2023-03
Informations de copyright
© 2024. The Author(s).
Références
Schaefer K, Erley CM, Herrath D, Stein G. Calcium salts of ketoacids as a new treatment strategy for uremic hyperphosphatemia. Kidney Int Suppl. 1989;27(5):136–9.
Schaefer K, Herrath D, Erley CM, Asmus G. Calcium ketovaline as new therapy for uremic hyperphosphatemia. Miner Electrolyte Metab. 1990;16(6):362–4.
Hao Y, Pan X, You J, Li G, Xu M, Rao Z. Microbial production of branched chain amino acids: advances and perspectives. Bioresour Technol. 2024;397:130502.
doi: 10.1016/j.biortech.2024.130502
Wang YY, Xu JZ, Zhang WG. Metabolic engineering of L-leucine production in Escherichia coli and Corynebacterium glutamicum: a review. Crit Rev Biotechnol. 2019;39(5):633–47.
doi: 10.1080/07388551.2019.1577214
Guo J, Sun X, Yuan Y, Chen Q, Ou Z, Deng Z, Ma T, Liu T. Metabolic engineering of Saccharomyces cerevisiae for vitamin B5 production. J Agric Food Chem. 2023;71(19):7408–17.
doi: 10.1021/acs.jafc.3c01082
Luo Z, Yu S, Zeng W, Zhou J. Comparative analysis of the chemical and biochemical synthesis of keto acids. Biotechnol Adv. 2021;47:107706.
doi: 10.1016/j.biotechadv.2021.107706
Zhou L, Zhu Y, Yuan Z, Liu G, Sun Z, Du S, Liu H, Li Y, Liu H, Zhou Z. Evaluation of metabolic engineering strategies on 2-ketoisovalerate production by Escherichia coli. Appl Environ Microbiol. 2022;88(17):e0097622.
doi: 10.1128/aem.00976-22
Carranza-Saavedra D, Torres-Bacete J, Blázquez B, Sánchez Henao CP, Zapata Montoya JE, Nogales J. System metabolic engineering of Escherichia coli W for the production of 2-ketoisovalerate using unconventional feedstock. Front Bioeng Biotechnol. 2023;11:1176445.
doi: 10.3389/fbioe.2023.1176445
Krause FS, Blombach B, Eikmanns BJ. Metabolic engineering of Corynebacterium glutamicum for 2-ketoisovalerate production. Appl Environ Microbiol. 2010;76(24):8053–61.
doi: 10.1128/AEM.01710-10
Buchholz J, Schwentner A, Brunnenkan B, Gabris C, Grimm S, Gerstmeir R, Takors R, Eikmanns BJ, Blombach B. Platform engineering of Corynebacterium glutamicum with reduced pyruvate dehydrogenase complex activity for improved production of L-lysine, L-valine, and 2-ketoisovalerate. Appl Environ Microbiol. 2013;79(18):5566–75.
doi: 10.1128/AEM.01741-13
Batianis C, van Rosmalen RP, Major M, van Ee C, Kasiotakis A, Weusthuis RA. Martins Dos Santos VAP. A tunable metabolic valve for precise growth control and increased product formation in Pseudomonas putida. Metab Eng. 2023;75:47–57.
doi: 10.1016/j.ymben.2022.10.002
Gu J, Zhou J, Zhang Z, Kim CH, Jiang B, Shi J, Hao J. Isobutanol and 2-ketoisovalerate production by Klebsiella pneumoniae via a native pathway. Metab Eng. 2017;43:71–84.
doi: 10.1016/j.ymben.2017.07.003
Pospíšil S, Kopecký J, Přikrylová V, Spížek J. Overproduction of 2-ketoisovalerate and monensin production by regulatory mutants of Streptomyces cinnamonensis resistant to 2-ketobutyrate and amino acids. FEMS Microbiol Lett. 1999;172(2):197–204.
doi: 10.1016/S0378-1097(99)00042-7
Cao M, Jiang T, Li P, Zhang Y, Guo S, Meng W, Lü C, Zhang W, Xu P, Gao C, Ma C. Pyruvate production from whey powder by metabolic engineered Klebsiella oxytoca. J Agric Food Chem. 2020;68(51):15275–83.
doi: 10.1021/acs.jafc.0c06724
Park JM, Song H, Lee HJ, Seung D. Genome-scale reconstruction and in silico analysis of Klebsiella oxytoca for 2,3-butanediol production. Microb Cell Fact. 2013;12(1):20.
doi: 10.1186/1475-2859-12-20
Wilson DJ. NIH guidelines for research involving recombinant DNA molecules. Account Res. 1993;3(2–3):177–85.
Xin B, Tao F, Wang Y, Liu H, Ma C, Xu P. Coordination of metabolic pathways: enhanced carbon conservation in 1,3-propanediol production by coupling with optically pure lactate biosynthesis. Metab Eng. 2017;41:102–14.
doi: 10.1016/j.ymben.2017.03.009
Khunnonkwao P, Jantama SS, Jantama K, Joannis-Cassan C, Taillandier P. Sequential coupling of enzymatic hydrolysis and fermentation platform for high yield and economical production of 2,3-butanediol from cassava by metabolically engineered Klebsiella oxytoca. J Chem Technol Biotechnol. 2021;96(5):1292–301.
doi: 10.1002/jctb.6643
In S, Khunnonkwao P, Wong N, Phosiran C, Jantama SS, Jantama K. Combining metabolic engineering and evolutionary adaptation in Klebsiella oxytoca KMS004 to significantly improve optically pure D-(-)-lactic acid yield and specific productivity in low nutrient medium. Appl Microbiol Biotechnol. 2020;104(22):9565–79.
doi: 10.1007/s00253-020-10933-0
Phosriran C, Wong N, Jantama K. An efficient production of bio-succinate in a novel metabolically engineered Klebsiella oxytoca by rational metabolic engineering and evolutionary adaptation. Bioresour Technol. 2024;393:130045.
doi: 10.1016/j.biortech.2023.130045
Cao M, Sun W, Wang S, Di H, Du Q, Tan X, Meng W, Kang Z, Liu Y, Xu P, Lü C, Ma C, Gao C. Efficient L-valine production using systematically metabolic engineered Klebsiella oxytoca. Bioresour Technol. 2024;395:130403.
doi: 10.1016/j.biortech.2024.130403
Macwan SR, Dabhi BK, Parmar SC, Aparnathi KD. Whey and its utilization. Int J Curr Microbiol App Sci. 2016;5(8):134–55.
doi: 10.20546/ijcmas.2016.508.016
Li P, Wang M, Di H, Du Q, Zhang Y, Tan X, Xu P, Gao C, Jiang T, Lü C, Ma C. Efficient production of 1,2,4-butanetriol from corn cob hydrolysate by metabolically engineered Escherichia coli. Microb Cell Fact. 2024;23(1):49.
doi: 10.1186/s12934-024-02317-0
Arslan NP, Aydogan MN, Taskin M. Citric acid production from partly deproteinized whey under non-sterile culture conditions using immobilized cells of lactose-positive and cold-adapted Yarrowia Lipolytica B9. J Biotechnol. 2016;231:32–9.
doi: 10.1016/j.jbiotec.2016.05.033
Mano J, Liu N, Hammond JH, Currie DH, Stephanopoulos G. Engineering Yarrowia Lipolytica for the utilization of acid whey. Metab Eng. 2020;57:43–50.
doi: 10.1016/j.ymben.2019.09.010
Novak K, Baar J, Freitag P, Pflügl S. Metabolic engineering of Escherichia coli W for isobutanol production on chemically defined medium and cheese whey as alternative raw material. J Ind Microbiol Biotechnol. 2020;47(12):1117–32.
doi: 10.1007/s10295-020-02319-y
Li Q, Sun B, Chen J, Zhang Y, Jiang Y, Yang S. A modified pCas/pTargetF system for CRISPR-Cas9-assisted genome editing in Escherichia coli. Acta Biochim Biophys Sin. 2021;53(5):620–7.
doi: 10.1093/abbs/gmab036
Gao C, Li Z, Zhang L, Wang C, Li K, Ma C, Xu P. An artificial enzymatic reaction cascade for a cell-free bio-system based on glycerol. Green Chem. 2015;17(2):804–7.
doi: 10.1039/C4GC01685H
McCully V, Burns G, Sokatch JR. Resolution of branched-chain oxo acid dehydrogenase complex of Pseudomonas aeruginosa PAO. Biochem J. 1986;233(3):737–42.
doi: 10.1042/bj2330737
Zhu K, Bayles DO, Xiong A, Jayaswal RK, Wilkinson BJ. Precursor and temperature modulation of fatty acid composition and growth of Listeria monocytogenes cold-sensitive mutants with transposon-interrupted branched-chain alpha-keto acid dehydrogenase. Microbiology. 2005;151(2):615–23.
doi: 10.1099/mic.0.27634-0
Lowe PN, Hodgson JA, Perham RN. Dual role of a single multienzyme complex in the oxidative decarboxylation of pyruvate and branched-chain 2-oxo acids in Bacillus subtilis. Biochem J. 1983;215(1):133–40.
doi: 10.1042/bj2150133
Singh VK, Hattangady DS, Giotis ES, Singh AK, Chamberlain NR, Stuart MK, Wilkinson BJ. Insertional inactivation of branched-chain alpha-keto acid dehydrogenase in Staphylococcus aureus leads to decreased branched-chain membrane fatty acid content and increased susceptibility to certain stresses. Appl Environ Microbiol. 2008;74(19):5882–90.
doi: 10.1128/AEM.00882-08
Hasegawa S, Uematsu K, Natsuma Y, Suda M, Hiraga K, Jojima T, Inui M, Yukawa H. Improvement of the redox balance increases L-valine production by Corynebacterium glutamicum under oxygen deprivation conditions. Appl Environ Microbiol. 2012;78(3):865–75.
doi: 10.1128/AEM.07056-11
Hao Y, Ma Q, Liu X, Fan X, Men J, Wu H, Jiang S, Tian D, Xiong B, Xie X. High-yield production of L-valine in engineered Escherichia coli by a novel two-stage fermentation. Metab Eng. 2020;62:198–206.
doi: 10.1016/j.ymben.2020.09.007
Prazeres AR, Carvalho F, Rivas J. Cheese whey management: a review. J Environ Manage. 2012;110:48–68.
doi: 10.1016/j.jenvman.2012.05.018
Kader Chowdhury QMM, Islam S, Narayanan L, Ogunleye SC, Wang S, Thu D, Freitag NE, Lawrence ML, Abdelhamed H. An insight into the role of branched-chain α-keto acid dehydrogenase (BKD) complex in branched-chain fatty acid biosynthesis and virulence of Listeria monocytogenes. J Bacteriol. 2024;206(7):e0003324.
doi: 10.1128/jb.00033-24
Mohedano MT, Konzock O, Chen Y. Strategies to increase tolerance and robustness of industrial microorganisms. Synth Syst Biotechnol. 2021;7(1):533–40.
doi: 10.1016/j.synbio.2021.12.009
Shen X, Wang J, Li C, Yuan Q, Yan Y. Dynamic gene expression engineering as a tool in pathway engineering. Curr Opin Biotechnol. 2019;59:122–9.
doi: 10.1016/j.copbio.2019.03.019
Buchanan D, Martindale W, Romeih E, Hebishy E. Recent advances in whey processing and valorisation: Technological and environmental perspectives. Int J Dairy Technol. 2023;76(2):291–312.
doi: 10.1111/1471-0307.12935
Tesfaw A, Oner ET, Assefa F. Evaluating crude whey for bioethanol production using non-Saccharomyces yeast, Kluyveromyces marxianus. SN Appl Sci. 2021;3(42):1–8.
Domínguez-Niño A, Cantú-Lozano D, Ragazzo-Sanchez JA, Andrade-González I, Luna-Solano G. Energy requirements and production cost of the spray drying process of cheese whey. Dry Technol. 2018;36(5):597–608.
doi: 10.1080/07373937.2017.1350863
Ma Y, Guo N, Wang S, Wang Y, Jiang Z, Guo L, Luo W, Wang Y. Metabolically engineer Clostridium saccharoperbutylacetonicum for comprehensive conversion of acid whey into valuable biofuels and biochemicals. Bioresour Technol. 2024;400:130640.
doi: 10.1016/j.biortech.2024.130640
Zhang K, Woodruff AP, Xiong M, Zhou J, Dhande YK. A synthetic metabolic pathway for production of the platform chemical isobutyric acid. ChemSusChem. 2011;4(8):1068–70.