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Production of LYZL6, a novel human c-type lysozyme, in recombinant Pichia pastoris employing high cell density fed-batch fermentation

Paper ID Volume ID Publish Year Pages File Format Full-Text
20289 43167 2014 6 PDF Available
Title
Production of LYZL6, a novel human c-type lysozyme, in recombinant Pichia pastoris employing high cell density fed-batch fermentation
Abstract

Lysozyme acts as an important defensive factor in innate immunity due to its well-recognized bacteriolytic activity. Here we describe the production and performance of human lysozyme-like 6 (LYZL6), a novel human c-type lysozyme homolog. A synthetic codon-optimized cDNA encoding the intact amino acid sequence of LYZL6 was cloned and expressed in Pichia pastoris SMD1168. Bioactive LYZL6 was successfully produced as a single major secreted protein with a molecular weight of 15 kDa, and exhibited bacteriolytic activity against Micrococcus lysodeikticus. The expression conditions were optimized, and the highest expression level of LYZL6 occurred when the recombinant strain was induced with 1.5% methanol under pH 4.5 at 24°C for 96 h. When high cell density fermentation of the recombinant P. pastoris was performed using a fed-batch strategy for totally 125 h in a 30 L fermenter, the dry cell weight and the extracellular lysozyme activity were increased to 116.3 g/L and 2340 U/mL, respectively. The LYZL6 protein concentration was 331 mg/L of fermentation supernatant, and the specific activity of LYZL6 towards M. lysodeikticus was 7069 U/mg. Therefore, we proved that LYZL6 is an antibacterial protein, suggesting a potential application of LYZL6 as an antimicrobial agent, and Pichia expression system for LYZL6 was successful and industrially promising.

Keywords
Lysozyme-like; Pichia pastoris; Expression optimization; Fed-batch fermentation; Bacteriolytic activity
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Production of LYZL6, a novel human c-type lysozyme, in recombinant Pichia pastoris employing high cell density fed-batch fermentation
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 118, Issue 4, October 2014, Pages 420–425
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us