Biosynthesis of non-animal chondroitin sulfate from methanol using genetically engineered Pichia pastoris

Abstract

Chondroitin sulfate (CS) is widely applied in the medical, clinical, and nutraceutical fields. However, all commercialized CS is extracted from animal tissues, which is associated with many problems, such as...

Heterologous production of chondroitin

“…The authors also optimized the fermentation process in bioreactor using K-12 which resulted in 27 μg/g dry cell weight of intracellular CS-A with a sulfation degree of 96%, a level of 4- O -sulfation similar to the ones found in animals. More recently, P. pastoris has been successfully engineered towards the production of CS-A using methanol as substrate [204] . The expression of kfoC and kfoA from E. coli K4, tuaD from B. subtilis , engineered mouse C4OST and overexpression of endogenous genes coding adenosine-5′-triphosphate sulfurylase (ATPS) and adenosine-5′-phosphosulfate kinase (APSK) to improve PAPS supply and sulfation, resulted in a production of 2.1 g/L CS-A with 4.0% sulfation using a fed-batch fermentation.…”

“…The expression of kfoC and kfoA from E. coli K4, tuaD from B. subtilis , engineered mouse C4OST and overexpression of endogenous genes coding adenosine-5′-triphosphate sulfurylase (ATPS) and adenosine-5′-phosphosulfate kinase (APSK) to improve PAPS supply and sulfation, resulted in a production of 2.1 g/L CS-A with 4.0% sulfation using a fed-batch fermentation. The integrated approach from these two recent studies [ 160 , 204 ] provides significant cost reduction to the process. However, the CS yields in E. coli are still low while the CS produced by engineered P. pastoris has a low sulfation degree.…”

Chondroitin Sulfate and Its Derivatives: A Review of Microbial and Other Production Methods

“…The most common strain used is E. coli K4 because it already contains the required chondroitin backbone. Both shake flasks and fermentation methods have been used to synthesize unsulfated chondroitin in E. coli K4 strain [61][62][63]. Most efforts to synthesize unsulfated chondroitin in E. coli K4 strain begin with deleting the chromosomal kfoE gene responsible for the fructosylation of chondroitin [57,64].…”

Microbiological-Chemical Sourced Chondroitin Sulfates Protect Neuroblastoma SH-SY5Y Cells against Oxidative Stress and Are Suitable for Hydrogel-Based Controlled Release

“…Microbial cell factories represent a promising approach. They can be engineered to produce CS directly [ 23 , 24 ] or combined with tailored chemical procedures aimed at decorating unsulfated chondroitin with well-defined sulfation patterns [ 25 ]. Noteworthy, the latter strategy leads to CS polysaccharides with biological responses comparable to animal sourced CS with the same sulfation degree [ 26 , 27 ].…”