Received: 18-05-2020
Accepted: 23-12-2020
DOI:
Views
Downloads
How to Cite:
The Effects of Cell Density, Temperature and pH on Lactoferrin Productionfrom Recombinant Pichia PastorisKM71H-3 Strain
,
Trinh Thi Thu Thuy
2, 3
Keywords
Lactoferrin, Pichia pastoris, KM71H-3, recombinant
Abstract
Lactoferrin (LF) is a glycoprotein of the transferrin family whose general function is to transport iron. LF is capable of enhancing the immune system, antibacterial, antioxidant, anti-tumor, and antiviral activities. The gene encoding LF derived from bovine has been successfully expressed in recombinant P. pastoris KM71H-3 strain on 2xMMP medium. This study aims at investigating the effects of culture conditions including initial biomass cell density (equivalent to OD600 value from 0.5-2.0), temperature (20°C, 24°C,28°C), and initial pH of the medium (5.5-7.5). The LF expression level was evaluated by SDS – PAGE and Dot bloting methods. The SDS- PAGE andDot - blot results showed that LF was expressed in 2xMMP mediumwell with the optimal conditions were at 28°C, pH 6.0 and the initial biomass cell density (OD600) 0.5.
References
Adlerova L., Bartoskova A. &Faldyna M. (2008). Lactoferrin_a review.Veterinarni Medicina.53(9): 457-468
Alamdari E., Niazi A., Yarizade A., MoghadamA. &Aram F. (2016). Expression of a Recombinant Therapeutic Protein, Lactoferrin, in PichiaPinkTM: a Powerful Antimicrobial Protein Biological Forum -An International Journal.8(1):471-478.
Barber M.F., KronenbergZ., Yandell M. &Elde N.C. (2016). Antimicrobial Functions of Lactoferrin Promote Genetic Conflicts in Ancient Primates and Modern Humans, PLoS Genet. 12(5): e1006063.
Bradford M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 7(72): 248-54.
Chahardooli M., Niazi A., AramF. &Sohrabi S.M. (2016). Expression of recombinant Arabian camel lactoferricin-related peptide in Pichia pastorisand its antimicrobial identification.J Sci Food Agric.96(2):569-75.
Chen H.L., LaiY.W., YenC.C., LinY.Y., LuC.Y., Yang S.H., Tsai T.C., Lin Y.J., LinC.W. &Chen C.M. (2004). Production of recombinant porcine lactoferrin exhibiting antibacterial activity in methylotrophic yeast, Pichia pastoris.J Mol Microbiol Biotechnol. 8(3): 141-9.
Gonzalez-Chavez S.A., Arevalo-Gallegos S. &Rascon-Cruz Q. (2009). Lactoferrin: structure, function and applications, Int J Antimicrob Agents.33(4): 301 e1-8.
Iglesias-Figueroa B., Valdiviezo-Godina N., Siqueiros-Cendon T., Sinagawa-GarciaS., Arevalo-GallegosS. &Rascon-Cruz Q. (2016). High-Level Expression of Recombinant Bovine Lactoferrin in Pichia pastoriswith Antimicrobial Activity. Int J Mol Sci. 17(6).
Nakamura I., Watanabe A., Tsunemitsu H., LeeN.Y., Kumura H., Shimazaki K.I. &Yagi Y. (2001). Production of recombinant bovine lactoferrin N-lobe in insect cells and its antimicrobial activity.Protein Expr Purif.21(3): 424-31.
Parkar D.R., JadhavR.N. &Pimpliskar M.R. (2015). Antibacterial Activity of Lactoferrin_A Review, Human journals.4(2):118-127.
Sharma D. (2015). Lactoferrin and Its Role in Neonatology: A Review Article, Journal of Pediatrics & Neonatal Care. 2(2).
Sharma R., Chakraborty D. &Gupta P. (2015). Bovine lactoferrin and its functions in animals -A review, Agricultural Reviews.36(4).
Thomassen E.a.J., VeenH.a.V., BerkelP.H.C.V., Nuijens J.H. &Abrahams J.P. (2005). The protein structure of recombinant human lactoferrin produced in the milk of transgenic cows closely matches the structure of human milk-derived lactoferrin, Transgenic Research. 14(4):397-405.
Trịnh Thị Thu Thủy, Nguyễn Thị Tình &Trương Quốc Phong (2018). Nghiên cứu tạo cấu trúc biểu hiện tái tổ hợp pPICZA::blfoptđể biểu hiện lactoferrin bò trong Pichia pastoris Kỷ yếu hội nghị công nghệ sinh học toàn quốc. tr.306-313.