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Food Chem ; 274: 305-313, 2019 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-30372943

RESUMO

The low expression level of acidic lipases from Aspergillus sp. remains a major obstacle for their use in industrial applications. In this study, fusion expression with three fusion partners was investigated to enhance the expression level of an acidic lipase from A. niger (ANL) in Pichia pastoris. When fused with a small ubiquitin-related modifier (SUMO), designated SANL, the highest activity reached 960 ±â€¯40 U/mL in a 3 L fermenter, which was 1.85-fold higher than that of the parent ANL. SANL exhibited its maximum activity at pH 2.5 and had lower Km and higher kcat/Km values than those of ANL. In gastrointestinal digestion experiments, SANL was resistant to pepsin and had high hydrolytic activity against triolein from pH 3.0 to 6.0. However, SANL was significantly inhibited by NaTDC above its CMC, which may limit its application for intestinal digestion, but allow it to remain suitable for gastric digestion.


Assuntos
Aspergillus niger/enzimologia , Lipase/genética , Pichia/genética , Proteínas Recombinantes de Fusão/genética , Aspergillus niger/genética , Celulose/metabolismo , Clonagem Molecular , Digestão , Suco Gástrico , Concentração de Íons de Hidrogênio , Hidrólise , Lipase/metabolismo , Microrganismos Geneticamente Modificados , Domínios Proteicos , Engenharia de Proteínas/instrumentação , Engenharia de Proteínas/métodos , Proteínas Recombinantes de Fusão/metabolismo
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