Abstract
Pichia pastoris, a methylotrophic yeast, is known to be an efficient host for heterologous proteins production. In this study, a recombinant P. pastoris Y11430 was found better for β-glucosidase activity in comparison with a wild type P. pastoris Y11430 strain, and thereby, subjected to methanol intermittent feed profiling for β-glucosidase production. The results showed that at 72 h of cultivation time, the cultures with 16.67% and 33.33% methanol feeding with constant rate could produce the total dry cell weight of 52.23 and 118.55 g/L, respectively, while the total mutant β-glucosidase activities were 1001.59 and 3259.82 units, respectively. The methanol feeding profile was kept at 33% with three methanol feeding strategies such as constant feed rate, linear feed rate, and exponential feed rate which were used in fed-batch fermentation. At 60 h of cultivation, the highest total mutant β-glucosidase activity was 2971.85 units for exponential feed rate culture. On the other hand, total mutant β-glucosidase activity of the constant feed rate culture and linear feed rate culture were 1682.25 and 1975.43 units, respectively. The kinetic parameters of exponential feed rate culture were specific growth rate on glycerol 0.228/h, specific growth of methanol 0.061/h, maximum total dry cell weight 196.73 g, yield coefficient biomass per methanol (\({Y_{x/m}}\)) 0.57 gcell/gMeOH, methanol consumption rate (\({Q_m}\)) 5.76 gMeOH/h, and enzyme productivity (\({Q_p}\)) 75.96 units/h. In conclusion, higher cell mass and β- glucosidase activity were produced under exponential feed rate than constant and linear feed rates.
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Acknowledgements
This research was supported by the Department of Biotechnology, Faculty of Agro-Industry, Kasetsart University, Thailand. The authors also acknowledge the support of Associate Professor Prachumporn Kongsaree, Department of Biochemistry, Kasetsart University, Bangkok, Thailand.
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Sirirat Changming: investigation and writing-original draft. Prachumporn Kongsaree: resource person and supervision. Imrana Niaz Sultan: validation and review and editing the draft. Afrasiab Khan Tareen: formal analysis, conceptualization, and review and editing draft. Wirat Vanichsriratana: project administration and resources. Sarote Sirisansaneeyakul: supervision and formal analysis. Pramuk Parakulsuksatid: conceptualization, supervision, project administration, and resources.
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Changming, S., Kongsaree, P., Sultan, I.N. et al. β-glucosidase production by recombinant Pichia pastoris strain Y1433 under optimal feed profiles of fed-batch cultivation. Folia Microbiol 68, 245–256 (2023). https://doi.org/10.1007/s12223-022-01008-w
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DOI: https://doi.org/10.1007/s12223-022-01008-w