Evaluation of Microbial Phospholipase and Lipase Activity Through the Chromatographic Analysis of Crude, Degummed, and Transesterified Soybean Oil for Biodiesel Production

J Sep Sci. 2024 Oct;47(19):e202400325. doi: 10.1002/jssc.202400325.

Abstract

The present study aimed at synthesizing fatty acid methyl esters in a combined enzymatic method by applying degumming and transesterification of soybean oil. A soluble lipase from Serratia sp. W3 and a recombinant phosphatidylcholine-preferring phospholipase C (PC-PLC) from Bacillus thuringiensis were used in a consecutive manner for phosphorus removal and conversion into methyl esters. By applying 1% of recombinant PC-PLC almost 83% of phosphorus was removed (final content of 21.01 mg/kg). Moreover, a sensitive and selective high-performance liquid chromatography method coupled to tandem mass spectrometry was applied to obtain a comprehensive lipid profile for the simultaneous evaluation of phospholipids removal and diacylglycerol (DAG) increase. A significant increase for all the monitored DAG species, up to 138.42%, was observed by using the enzymatic degumming, in comparison to the crude sample, resulting in an increased oil yield. Serratia sp. W3 lipase was identified as a suitable biocatalyst for biodiesel production, converting efficiently the acylglycerols. The results regarding the physical-chemical characteristics show that the cetane level, density and pour point of the obtained biodiesel are close to current regulation requirements. These findings highlight the potential of a two-step process implementation, based on the combination of lipase and phospholipase, as a suitable alternative for biodiesel production.

Keywords: Bacillus thuringiensis phosphatidylcholine‐preferring phospholipase C; Serratia sp W3 lipase; diacylglycerols and phosphorus content; enzymatic degumming; vegetable oil transesterification.

MeSH terms

  • Bacillus thuringiensis / chemistry
  • Bacillus thuringiensis / enzymology
  • Bacillus thuringiensis / metabolism
  • Biofuels* / analysis
  • Chromatography, High Pressure Liquid
  • Esterification
  • Lipase* / chemistry
  • Lipase* / metabolism
  • Phospholipases / chemistry
  • Phospholipases / metabolism
  • Serratia* / chemistry
  • Serratia* / enzymology
  • Serratia* / metabolism
  • Soybean Oil* / chemistry
  • Soybean Oil* / metabolism
  • Tandem Mass Spectrometry

Substances

  • Lipase
  • Biofuels
  • Soybean Oil
  • Phospholipases