Investigation of Structural Features of Two Related Lipases and the Impact on Fatty Acid Specificity in Vegetable Fats

Dong, Zehui; Olofsson, Kim; Linares-Pastén, Javier A.; Karlsson, Eva Nordberg

doi:10.3390/ijms23137072

Cited by 3 publications

(1 citation statement)

References 36 publications

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“…Lipases from filamentous fungi families, such as Rhizopus oryzae (ROL), have been widely used in the oil and fats industry, due to their high 1,3-regioselectivity toward triglycerides that make them versatile in lipid modification (Rodrigues et al 2013 ). The structural features of one of the major native lipases from Y. lipolytica , Lip2 (Bordes et al 2010 ), is very similar to ROL (Dong et al 2022 ), making Y. lipolytica a potential excellent host for ROL production.…”

Section: Introductionmentioning

confidence: 99%

Production of Rhizopus oryzae lipase using optimized Yarrowia lipolytica expression system

Vidal

Dong

Olofsson³

et al. 2023

FEMS Yeast Research

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Yarrowia lipolytica is an alternative yeast for heterologous protein production. Based on auto-cloning vectors, a set of 18 chromogenic cloning vectors was developed, each containing one of the excisable auxotrophic selective markers URA3ex, LYS5ex and LEU2ex, and one of six different promoters: the constitutive pTEF, the phase dependent hybrid pHp4d, and the erythritol-inducible promoters from pEYK1 and pEYL1 derivatives. These vectors allowed to increase the speed of cloning of the gene of interest. In parallel, an improved new rProt recipient strain JMY8647 was developed by abolishing filamentation and introducing an auxotrophy for lysine (Lys−), providing an additional marker for genetic engineering. Using this cloning strategy, the optimal targeting sequence for Rhizopus oryzae ROL lipase secretion was determined. Among the eight targeting sequences, the SP6 signal sequence resulted in a 23% improvement in the lipase activity compared to that obtained with the wild-type ROL signal sequence. Higher specific lipase activities were obtained using hybrid erythritol-inducible promoters pHU8EYK and pEYL1-5AB, 1.9 and 2.2 times respectively, when compared with the constitutive pTEF promoter. Two copy strains produce a 3.3 fold increase in lipase activity over the pTEF monocopy strain (266.7 mU/mg versus 79.7 mU/mg).

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Section: Introductionmentioning

confidence: 99%

Production of Rhizopus oryzae lipase using optimized Yarrowia lipolytica expression system

Vidal

Dong

Olofsson³

et al. 2023

FEMS Yeast Research

Self Cite

View full text Add to dashboard Cite

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Transcriptome Sequencing Reveals Key Genes for Sunflower Unsaturated Fatty Acid Synthesis

et al. 2023

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Sunflower (Helianthus annuus L.) is an important oil crop with rich nutrients, and genetically engineered breeding has become an important way to improve its quality. In this study, five varieties of oilseed sunflower were analyzed for fatty acid (FA) content. The seed embryos of one of the high oleic acid (OA) varieties were transcriptome sequenced at different stages. The results showed that OA synthesis dominated the unsaturated fatty acid (UFA) synthesis pathways in seed embryos. Substantially differentially expressed genes were detected at various post-flowering stages. Specifically, the up-regulated gene numbers were highest at 10 d after flowering, while most genes were down-regulated at 20 d after flowering. The enriched genes were rather consistent with almost all experimental groups exhibiting enrichment to the FAD2 gene. The expression of FAD2 was highly negatively correlated with the expressions of FAD6, FAD3, and FAD7. During seed embryo development, the expression level of FAD2 was highly negatively correlated with the final OA content and was highly positively correlated with the final linoleic acid (LA) content. This suggests that the FAD2 is a key enzyme catalyzing the OA to LA conversion.

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The Effects of Buffer Nature on Immobilized Lipase Stability Depend on Enzyme Support Loading

Abellanas-Perez,

Carballares,

Rocha-Martin

et al. 2024

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The lipases from Thermomyces lanuginosus (TLL) and Candida antarctica (B) (CALB) were immobilized on octyl-agarose beads at 1 mg/g (a loading under the capacity of the support) and by overloading the support with the enzymes. These biocatalysts were compared in their stabilities in 10 mM of sodium phosphate, HEPES, and Tris-HCl at pH 7. Lowly loaded CALB was more stable than highly loaded CALB preparation, while with TLL this effect was smaller. Phosphate was very negative for the stability of the CALB biocatalyst and moderately negative using TLL at both loadings. The stability of the enzymes in HEPES and Tris-HCl presented a different response as a function of the enzyme loading (e.g., using lowly loaded CALB, the stabilities were similar in both buffers, but it was clearly smaller in HEPES using the highly loaded biocatalysts). Moreover, the specific activity of the immobilized enzymes versus p-nitrophenol butyrate, triacetin and R- or S-methyl mandelate depended on the buffer, enzyme loading, and interaction between them. In some cases, almost twice the expected activity could be obtained using highly loaded octyl-CALB, depending on the buffer. A co-interaction between the effects on enzyme activity and the specificity of support enzyme loading and buffer nature was detected.

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Investigation of Structural Features of Two Related Lipases and the Impact on Fatty Acid Specificity in Vegetable Fats

Cited by 3 publications

References 36 publications

Production of Rhizopus oryzae lipase using optimized Yarrowia lipolytica expression system

Production of Rhizopus oryzae lipase using optimized Yarrowia lipolytica expression system

Transcriptome Sequencing Reveals Key Genes for Sunflower Unsaturated Fatty Acid Synthesis

The Effects of Buffer Nature on Immobilized Lipase Stability Depend on Enzyme Support Loading

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