Ceramide is a class of lipid molecules used in cosmetics and pharmaceuticals.The synthesis of ceramide by microbial metabolic engineering has important prospects.Pichia pastoris is an excellent industrial host.At present, there is no report on the use of P.pastoris to enhance synthesis of ceramide.Firstly, P.pastoris GS115ΔPAS_chr3_0329 (G-K) was constructed as the chassis, and then the ceramide synthesis negative feedback gene PAS_chr4_0427 was knocked out in G-K to obtain G-KO, and the composition and content of ceramide were investigated by lipidomics.Results showed that compared with G-K, the relative content of ceramide in total lipid in G-KO increased by 628.13% to 9.32%.Moreover, the deletion of gene PAS_chr4_0427 promoted the synthesis of more ceramides with unsaturated long-chain bases.At the same time, the relative content of sphingolipids in G-KO increased, while the relative content of phospholipids decreased.HPLC analysis of Cer(d18∶0) ceramide with the highest ceramide abundance in P.pastoris showed that the content of Cer(d18∶0) ceramide in G-KO reached 90.22 mg/L, which was 872.20% higher than that of G-K.This study confirmed that P.pastoris could be used for ceramide synthesis.The PAS_chr4_0427 deletion strain G-KO has excellent ceramide synthesis ability and has the potential for industrial fermentation to prepare ceramide.
HUANG Ming
,
WU Jiaxin
,
ZHANG Mu
,
WANG Shuiping
,
HU Xiaoqing
. Knockout of PAS_chr4_0427 promotes ceramide synthesis in Pichia pastoris[J]. Food and Fermentation Industries, 2024
, 50(17)
: 17
-22
.
DOI: 10.13995/j.cnki.11-1802/ts.039574
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