The composition of the carbon source and the time of cell harvest are critical determinants of the final yield of human papillomavirus type 16 L1 protein produced in Saccharomyces cerevisiae

Kim, Hyoung Jin; Kwag, Hye-Lim; Jin, Yingji; Kim, Hong-Jin

doi:10.1016/j.pep.2011.06.010

Cited by 10 publications

(10 citation statements)

References 38 publications

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“…Previously we found that the levels of conformational neutralizing epitopes on the surface of HPV16 VLPs increased with increasing culture duration when the culture contained more than 6% carbon source [17]. In the same study, we found that the proportion of soluble L1 protein in culture increased with longer culture duration [17].…”

Section: Discussionsupporting

confidence: 58%

“…In the same study, we found that the proportion of soluble L1 protein in culture increased with longer culture duration [17]. These results demonstrated that in the yeast expression system longer culture periods may be necessary to obtain correctly folded L1 proteins that go on to form highly immunogenic HPV L1 VLPs.…”

Section: Discussionmentioning

confidence: 56%

“…In this study, we compared the antigenicity and immunogenicity of HPV type 16 L1 protein (HPV16 L1) VLPs produced in the S. cerevisiae expression system with different concentrations of carbon source. Previously, we found that the concentration and composition of the carbon source used for S. cerevisiae producing HPV16 L1 protein significantly affect the yield of the HPV16 L1 protein [17]. In this study, we report that the concentration of the carbon source in yeast cultures substantially affects the quality of HPV16 L1 VLPs.…”

Section: Introductionmentioning

confidence: 67%

“…Cells were culture in YPDG medium containing 7% glucose and 1% galactose for 144 h at 30°C. This culture condition showed the highest production yield of the L1 protein (see reference [17]). The intracellular levels of GSSG and GSH were determined as described [31] with modification.…”

Section: Supporting Informationmentioning

confidence: 87%

“…SDS-PAGE and Western blotting were performed as described [17]. Tubulin was detected using rat anti-tubulin antibody (Abcam, USA).…”

Section: Methodsmentioning

confidence: 99%

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The Concentration of Carbon Source in the Medium Affects the Quality of Virus-Like Particles of Human Papillomavirus Type 16 Produced in Saccharomyces cerevisiae

2014

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There is accumulating evidence that virus-like particles (VLPs) recombinantly produced in Saccharomyces cerevisiae (S. cerevisiae) are characterized by low structural stability, and that this is associated with reduced antigenicity and immunogenicity. However, little attention has been devoted to methods of improving the quality of the VLPs. Here, we investigated the effect of carbon source concentration in the medium on the antigenicity and immunogenicity of human papillomavirus (HPV) type 16 L1 VLPs expressed in S. cerevisiae from the galactose promoter. Media containing 2, 4, 6, and 8% carbon source, composed of both glucose and galactose in equal proportion, were used. VLP antigenicity was enhanced in cultures grown on media with 6 or 8% carbon source, compared to those from cultures with less than 6% carbon source. Moreover, the VLPs obtained from these cultures induced higher anti-HPV16 L1 IgG titers and neutralizing antibody titers in immunized mice than those purified from cultures with less than 6% carbon source. Our results indicate that the concentration of the carbon source in the medium plays a crucial role in determining the antigenicity and immunogenicity of HPV type16 L1 VLPs.

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Section: Discussionsupporting

confidence: 58%

Section: Discussionmentioning

confidence: 56%

Section: Introductionmentioning

confidence: 67%

Section: Supporting Informationmentioning

confidence: 87%

“…SDS-PAGE and Western blotting were performed as described [17]. Tubulin was detected using rat anti-tubulin antibody (Abcam, USA).…”

Section: Methodsmentioning

confidence: 99%

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The Concentration of Carbon Source in the Medium Affects the Quality of Virus-Like Particles of Human Papillomavirus Type 16 Produced in Saccharomyces cerevisiae

2014

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Codon optimization of the human papillomavirus type 58 L1 gene enhances the expression of soluble L1 protein in Saccharomyces cerevisiae

2012

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The effect of codon optimization of L1 gene on the production of the L1 protein of human papillomavirus (HPV) was investigated in a yeast expression system. Saccharomyces cerevisiae was transformed with a plasmid containing either the wild type (WS)-HPV type 58 L1 (HPV58 L1) gene or a codon-optimized (MO)-HPV58 L1 gene. The proportion of soluble L1 protein expressed from MO-HPV58 L1 was significantly higher than that expressed from WS-HPV58 L1. Moreover, the amount of purified MO-HPV58 L1 protein recovered was 2.5-fold higher than the amount of WS-HPV58 L1 protein. Codon optimization of HPV58 L1 gene thus increases the proportion of soluble L1 protein and the amount of purified product that can be used as antigen to generate vaccines.

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Diverse expression levels of two codon-optimized genes that encode human papilloma virus type 16 major protein L1 in Hansenula polymorpha

et al. 2014

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Two versions of an optimized gene that encodes human papilloma virus type 16 major protein L1 were designed according to the codon usage frequency of Pichia pastoris. Y16 was highly expressed in both P. pastoris and Hansenula polymorpha. M16 expression was as efficient as that of Y16 in P. pastoris, but merely detectable in H. polymorpha even though transcription levels of M16 and Y16 were similar. H. polymorpha had a unique codon usage frequency that contains many more rare codons than Saccharomyces cerevisiae or P. pastoris. These findings indicate that even codon-optimized genes that are expressed well in S. cerevisiae and P. pastoris may be inefficiently expressed in H. polymorpha; thus rare codons must be avoided when universal optimized gene versions are designed to facilitate expression in a variety of yeast expression systems, especially H. polymorpha is involved.

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The composition of the carbon source and the time of cell harvest are critical determinants of the final yield of human papillomavirus type 16 L1 protein produced in Saccharomyces cerevisiae

Cited by 10 publications

References 38 publications

The Concentration of Carbon Source in the Medium Affects the Quality of Virus-Like Particles of Human Papillomavirus Type 16 Produced in Saccharomyces cerevisiae

The Concentration of Carbon Source in the Medium Affects the Quality of Virus-Like Particles of Human Papillomavirus Type 16 Produced in Saccharomyces cerevisiae

Codon optimization of the human papillomavirus type 58 L1 gene enhances the expression of soluble L1 protein in Saccharomyces cerevisiae

Diverse expression levels of two codon-optimized genes that encode human papilloma virus type 16 major protein L1 in Hansenula polymorpha

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