ArticlePloS one2016
A Simplified and Efficient Process for Insulin Production in Pichia pastoris.
Article in PloS one, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- Impact of Ethanol Stress onFoods (Basel, Switzerland) · 2025Article
- Balancing Doses of EL222 and Light Improves Optogenetic Induction of Protein Production in Komagataella phaffii.Biotechnology and bioengineering · 2025Article
- Review
- A simple and effective method to remove pigments from heterologous secretory proteins expressed in Pichia pastoris.Advanced biotechnology · 2024Article
- Large-scale crystallization as an intermediate processing step in insulin downstream process: explored advantages and identified tool for process intensification.Bioprocess and biosystems engineering · 2023Article
- Understanding Biosimilar Insulins - Development, Manufacturing, and Clinical Trials.Journal of diabetes science and technology · 2023Review
- Advances in Metabolic Engineering ofJournal of fungi (Basel, Switzerland) · 2023Review
- Full-length versus truncated α-factor secretory signal sequences for expression of recombinant human insulin precursor in yeast Pichia pastoris: a comparison.Journal, genetic engineering & biotechnology · 2023Article
- Semisynthesis of Aminomethyl-Insulin: An Atom-Economic Strategy to Increase the Affinity and Selectivity of a Protein for Recognition by a Synthetic Receptor.Bioconjugate chemistry · 2023Article
- Current advances and future prospects in production of recombinant insulin and other proteins to treat diabetes mellitus.Biotechnology letters · 2022Review
- Buried Kex2 Sites in Glargine Precursor Aggregates Prevent Its Intracellular Processing in Pichia pastoris MutApplied biochemistry and biotechnology · 2021Article
- An effective combination of codon optimization, gene dosage, and process optimization for high-level production of fibrinolytic enzyme in Komagataella phaffii (Pichia pastoris).BMC biotechnology · 2020Article
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Authors and funding
7 authors.
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No grant is acknowledged in the PubMed record.
Abstract
A significant barrier to insulin is affordability. In this manuscript we describe improvements to key steps in the insulin production process in Pichia pastoris that reduce cost and time. The strategy for recovery and processing of human insulin precursor has been streamlined to two steps from bioreactor to the transpeptidation reaction. In the first step the insulin precursor secreted during the methanol induction phase is recovered directly from the culture broth using Tangential Flow Filtration with a Prostak™ module eliminating the laborious and time-consuming multi-step clarification, including centrifugation. In the second step the protein is applied at very high loadings on a cation exchange resin and eluted in a mixture of water and ethanol to obtain a concentrated insulin precursor, suitable for use directly in the transpeptidation reaction. Overall the yield from insulin precursor to human insulin was 51% and consisted of three purification chromatography steps. In addition we describe a method for recovery of the excess of H-Thr(tBu)-OtBu from the transpeptidation reaction mixture, one of the more costly reagents in the process, along with its successful reuse.
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