ArticleJournal of industrial microbiology & biotechnology2014
Enhanced production of Thermomyces lanuginosus lipase in Pichia pastoris via genetic and fermentation strategies.
Article in Journal of industrial microbiology & biotechnology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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10 citing papers in PubMed, 39 citations in OpenAlex.
- Article
- Production and Functional Evaluation of Recombinant Active Peptide RH in Pichia Pastoris: Protection Against Escherichia Coli Induced Cell Death.Probiotics and antimicrobial proteins · 2026Article
- Employing Engineered Enolase Promoter for Efficient Expression ofInternational journal of molecular sciences · 2022Article
- 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
- Hybrid-architectured double-promoter expression systems enhance and upregulate-deregulated gene expressions in Pichia pastoris in methanol-free media.Applied microbiology and biotechnology · 2020Article
- Development of a New High-Cell Density Fermentation Strategy for Enhanced Production of a Fungus β-Glucosidase inFrontiers in microbiology · 2020Article
- Enhancement of thermoalkaliphilic xylanase production by Pichia pastoris through novel fed-batch strategy in high cell-density fermentation.BMC biotechnology · 2017Article
- Chitinase from Thermomyces lanuginosus SSBP and its biotechnological applications.Extremophiles : life under extreme conditions · 2015Review
- Bioprocess for efficient production of recombinant Pichia anomala phytase and its applicability in dephytinizing chick feed and whole wheat flat Indian breads.Journal of industrial microbiology & biotechnology · 2015Article
- Beauveria bassiana Lipase A expressed in Komagataella (Pichia) pastoris with potential for biodiesel catalysis.Frontiers in microbiology · 2015Article
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6 authors at 1 institution in 1 country.
Funding
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Abstract
This study attempted to enhance the expression level of Thermomyces lanuginosus lipase (TLL) in Pichia pastoris using a series of strategies. The tll gene was first inserted into the expression vector pPIC9 K and transformed into P. pastoris strain GS115. The maximum hydrolytic activity of TLL reached 4,350 U/mL under the optimal culture conditions of a 500 mL shaking flask containing 20 mL culture medium with the addition of 1.2 % (w/v) methanol, cultivation for 144 h at pH 7.0 and 27 °C. To further increase the TLL expression and copy number, strains containing two plasmids were obtained by sequential electroporation into GS115/9k-TLL #3 with a second vector, either pGAPZαA-TLL, pFZα-TLL, or pPICZαA-TLL. The maximum activity of the resultant strains GS115/9KTLL-ZαATLL #40, GS115/9KTLL-FZαATLL #46 and GS115/9KTLL-GAPTLL #45 was 6,600 U/mL, 6,000 U/mL and 4,800 U/mL, respectively. The tll copy number in these strains, as assessed by real-time quantitative PCR, was demonstrated to be seven, five, and three, respectively, versus two copies in GS115/9k-TLL #3. When a co-feeding strategy of sorbitol/methanol was adopted in a 3-L fermenter, the maximum TLL activity of GS115/9k-TLL #3 increased to 27,000 U/mL after 130 h of fed-batch fermentation, whereas, the maximum TLL activity was 19,500 U/mL after 145 h incubation when methanol was used as the sole carbon source.
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