Heterologous Expression of Biologically Active Porcine Lactoferrin in Pichia Pastoris Yeast

Pecorini, C.; Savazzini, F.; Martino, Piera Anna; Fusi, E.; Fogher, Corrado; Baldi, Alfonso

doi:10.1007/s11259-005-0086-1

Cited by 12 publications

(6 citation statements)

References 11 publications

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“…NicaTf was expressed in P. pastoris cells. This system has been used to express other transferrins (29)(30)(31). The purification is illustrated in SI Fig.…”

Section: Resultsmentioning

confidence: 99%

On the evolutionary significance and metal-binding characteristics of a monolobal transferrin fromCiona intestinalis

Tinoco

Peterson

Lucchese

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

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“…NicaTf was expressed in P. pastoris cells. This system has been used to express other transferrins (29)(30)(31). The purification is illustrated in SI Fig.…”

Section: Resultsmentioning

confidence: 99%

On the evolutionary significance and metal-binding characteristics of a monolobal transferrin fromCiona intestinalis

Tinoco

Peterson

Lucchese

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

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“…The Pichia pastoris system has now been used successfully to express a large number of intracellular and secreted proteins. Extremely high levels have been obtained for some biologically active molecules [6], and LF has been shown to be expressed in P. pastoris with the highest yield of 115 mg·L −1 [17,18,25,28]. In this paper, the effect of the ratio of glycerol and methanol on the expression of hLF and the growth of yeast cells were determined by dot blotting and OD600.…”

Section: Optimization Of Hlf Expressionmentioning

confidence: 99%

High-level expression and production of human lactoferrin in Pichia pastoris

Jiang

Chen

Jia

et al. 2008

Dairy Sci. Technol.

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-Lactoferrin (LF) is a multifunctional iron-binding glycoprotein which is found in high concentrations in milk. Recombinant human LF (rhLF) may provide health benefits. In the present study, we report an optimization of the production system of recombinant human lactoferrin that has enabled the production of recombinant protein at levels up to 1200 mg·L −1 . The hLF was expressed in the methylotrophic yeast Pichia pastoris using the pPIC9K vector. The expression level of recombinant hLF (rhLF) was improved significantly by mixed methanol/glycerol feeding at the ratio of 4:1 during the induction phase of high cell-density fermentation. A yield of approximately 1200 mg·L −1 was obtained in fed-batch fermentation with this system, which was much higher than the reported values for other systems (1 mg·L −1 to 115 mg·L −1 ). The rhLF was purified via ionexchange chromatography using SP Sepharose TM Fast Flow and has a similar molecular mass of 80 kg·mol −1 to native hLF. The level of glycosylation of the recombinant protein is similar to that of the native protein.human lactoferrin / Pichia pastoris / high cell-density fermentation / glycosylationRésumé -Haut niveau d'expression et de production de lactoferrine humaine par Pichia pastoris. La lactoferrine (LF) est une glycoprotéine multifonctionnelle liant le fer, qui se trouve en concentration élevée dans le lait. La lactoferrine humaine recombinante (rhLF) peut être bénéfique pour la santé. Dans cette étude, nous présentons une optimisation du système de production de rhLF permettant d'obtenir des niveaux de production élevés. La lactoferrine humaine était exprimée dans la levure méthylotrophe Pichia pastoris à l'aide du vecteur pPIC9K. Le niveau d'expression de rhLF était amélioré significativement lorsque le milieu était alimenté d'un mélange méthanol/glycérol dans un ratio de 4:1 pendant la phase d'induction de la fermentation à haute densité cellulaire.

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“…Moreover, mice fed milk supplemented with bovine lactoferrin had a reduced proliferation of Clostridium species in the gut (Teraguchi et al 1995). Pecorini et al (2003) tested the antimicrobial activity of recombinant porcine lactoferrin, expressed in Pichia pastoris yeast, on Esch. coli strains, isolated from piglets with neonatal and weaning diarrhoea.…”

Section: Lactoferrin -Milk Bioactive Component Modulating Intestinal mentioning

confidence: 99%

Biological effects of milk proteins and their peptides with emphasis on those related to the gastrointestinal ecosystem

Baldi

Politis

Pecorini

et al. 2005

Journal of Dairy Research

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The protein fraction of milk contains several components with physiological significance for the development of the newborn. Among them, immunomodulatory peptides and lactoferrin exemplify the complexity of biologically active substances of milk. Immunomodulatory peptides have latent activity within the native protein and are generated after proteolysis during gastrointestinal transit. Once they are generated, they modulate mucosal immunity, possibly by guiding the local immune system until it develops its full functionality. Lactoferrin is another milk bioactive compound with nutritional and health promoting properties; it modulates the microbial intestinal environment, displays anti-microbial activity against various pathogens and stimulates the establishment of beneficial microflora. The following overview focuses on the importance of immunomodulatory peptides and lactoferrin for the maturation of intestine and immune system that are functionally immature in the newborn.

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Heterologous Expression of Biologically Active Porcine Lactoferrin in Pichia Pastoris Yeast

Cited by 12 publications

References 11 publications

On the evolutionary significance and metal-binding characteristics of a monolobal transferrin fromCiona intestinalis

On the evolutionary significance and metal-binding characteristics of a monolobal transferrin fromCiona intestinalis

High-level expression and production of human lactoferrin in Pichia pastoris

Biological effects of milk proteins and their peptides with emphasis on those related to the gastrointestinal ecosystem

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