High-level secretory production of recombinant human platelet-derived growth factor-BB by Saccharomyces cerevisiae under the non-selective conditions

Wang, Y.; Xue, Lu‐Lu; Li, Y.; Zhu, Yan; Yang, Bo; Wang, X.

doi:10.1134/s0003683809020070

Cited by 3 publications

(4 citation statements)

References 16 publications

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“…Từ chủng nấm men này, Dương Long Duy và cộng sự (2015) đã lên men sản xuất thành công protein rhPDGF-BB ở quy mô 0,5 L, sử dụng methanol vừa làm nguồn carbon, vừa làm chất cảm ứng trong giai đoạn biểu hiện protein với sản lượng tiết protein mục tiêu đạt 686,69 mg/L, chiếm 57,02% tổng lượng protein tiết 4 . Đây là năng suất sản xuất tiềm năng, cao hơn các công bố sử dụng các chủng chủ khác nhau trước đây trên thế giới 5,6 . Tuy nhiên, việc sử dụng hoàn toàn methanol trong giai đoạn cảm ứng, nhất là ở quy mô lớn khi hiệu quả khuấy trộn bị giảm sút, có thể gây ức chế biến dưỡng và sản xuất protein của chủng nấm men tái tổ hợp.…”

Section: Giới Thiệuunclassified

Study of glycerol-methanol and sorbitol-methanol mixed feed strategies for recombinant hPDGF-BB production in Pichia pastoris

Trần

Lê²,

Nguyễn³

et al. 2021

Sci.Tech.Dev.J.-Eng.Tech.

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The recombinant human platelet-derived growth factor type BB (rhPDGF-BB) protein has been approved by FDA for diabetic foot ulcer treatment due to its function of stimulating cell proliferation and wound healing. In this study, the production of rhPDGF-BB from Pichia pastoris is studied in a 7.5 L fermentation using methanol as carbon source and inducer in induction phase. The specific growth rate of the recombinant strain reduced 1.833-fold and rhPDGF-BB productivity reduced 2.13-fold compared to at 0.5 L. To improve growth and protein expression of the strain in induction phase, the mixed feed strategy, in which glycerol or sorbitol was co-fed with methanol at three different molar ratios (1:1, 1:2 and 1:3) was conducted. Using the mixture of glycerol and methanol at the molar ratio 1:2 and the mixture of sorbitol and methanol at the molar ratio 1:3 increased the productivity of rhPDGF-BB protein 1.32-fold (280,48 mg/L) and 1.24-fold (264,15 mg/L), respectively, higher than using only methanol in the induction phase of the fermentation process.

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Section: Giới Thiệuunclassified

Study of glycerol-methanol and sorbitol-methanol mixed feed strategies for recombinant hPDGF-BB production in Pichia pastoris

Trần

Lê²,

Nguyễn³

et al. 2021

Sci.Tech.Dev.J.-Eng.Tech.

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“…The protocol was optimized from the study of Wang et al [7] . Cells were separated from the biomass by centrifugation at 6000 rpm, 4°C.…”

Section: Purification Of Rhpdgf-bb-mentioning

confidence: 99%

“…The rhPDGF-BB has been approved by FDA as the treatment for diabetic foot ulcer and self-bone grafting [1] . It has been produced in a variety of heterologous systems including Escherichia coli [2][3][4] , Chinese hamster ovary cells [5] , Saccharomyces cerevisiae [6,7] , baculovirus [8] , vaccinia viruses [9] , mushroom [10] , plant [11] and Pichia pastoris [12] . Babavalian et al [12] reported a high efficiency of 30 mg/L in Pichia pink, a P. pastoris mutant cell, without optimization.…”

Section: Introductionmentioning

confidence: 99%

Storage of the Recombinant Protein hPDGF-BB in the Culture of Pichia pastoris

Pham¹,

Quoc²,

Le³

et al. 2018

IJLSSR

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Human platelet-derived growth factor-BB (hPDGF-BB), a proliferation factor, has been successfully manufactured and approved by FDA as the treatment for diabetic foot ulcer and self bone grafting. There have been no reports on the storage of the recombinant hPDGF-BB (rhPDGF-BB) in the Pichia pastoris fermentation broth although during the research of process development and the production manufacture it needs to be stored at low temperature. The concentration of rhPDGF-BB protein in the fed-batch fermentation broth of P. pastoris was stable during 3-week storage at -20 o C, but its bioactivity was reduced by 20%. The addition of a mixture of 50% glycerol with either 1 mM EDTA or 1 mM PMSF into the fermentation broth could fully preserve the bioactivity of rhPDGF-BB until 3 weeks at -20 o C. The addition of 50% glycerol with either 1 mM EDTA or 1 mM PMSF was found no affection on the protein purification process.

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“…Thú vị hơn nữa, khi chúng tôi tiến hành điện di protein thu được từ nấm men trên gel polyacrylamide không bổ sung tác nhân biến tính (hình 6b), chúng tôi nhận thấy protein tiết thu được ở các chủng nấm men có kích thước khoảng 31 kDa. Theo các nghiên cứu của Raines (1982) [8] và Wang (2008) [9] đây có thể là trạng thái dimer của protein PDGF. Điều này cho thấy khả năng thu nhận protein PDGF ở trạng thái dimer (là trạng thái có hoạt tính sinh học) từ các chủng nẩm men tái tổ hợp.…”

Section: Tạo Dòng Tế Bào P Pastoris X33 Mang Gen Mã Hóa Pdgfunclassified

A study on constructing and screening of multi-copy pdgf recombinant Pichia pastoris clones producing high yield of PDGF-BB (platelet derived growth factor BB)

Khánh¹,

Trang²,

Thanh³

et al. 2014

V J Bio

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Platelet-derived growth factor-BB has been approved by US Food and Drug Administration-FDA for the treatment of neuropathic ulcers. Since the number of diabetic patients has increased rapidly, it is necessary to study the production of PDGF-BB with low cost for treatment. Among the expression systems used for human recombinant protein expression, Pichia pastoris is the remarkable one with many advantages. Multiple integrated copies of target gene may give advance in driving up expression level of target protein. In this study, we succeeded in constructing and screening two yeast clones, which carry multi-copy of pdgf target gene. The two multi-copy yeast clones were subjected indetermination of PDGF expression level and compared to single-copy yeast clones. Our results showed that multi-copy pdgf yeast clones have higher level expression of target protein. PDGF produced by the multi-copy pdgf yeast clones accounted for over 50% of total secretory protein, and was folded in dimer form, which is necessary for performing biofunction in wound healing.

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High-level secretory production of recombinant human platelet-derived growth factor-BB by Saccharomyces cerevisiae under the non-selective conditions

Cited by 3 publications

References 16 publications

Study of glycerol-methanol and sorbitol-methanol mixed feed strategies for recombinant hPDGF-BB production in Pichia pastoris

Study of glycerol-methanol and sorbitol-methanol mixed feed strategies for recombinant hPDGF-BB production in Pichia pastoris

Storage of the Recombinant Protein hPDGF-BB in the Culture of Pichia pastoris

A study on constructing and screening of multi-copy pdgf recombinant Pichia pastoris clones producing high yield of PDGF-BB (platelet derived growth factor BB)

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