Comparison of the biochemical properties of a recombinant lipase extract from Rhizopus oryzae expressed in Pichia pastoris with a native extract

2011 ◽  
Vol 54 (2) ◽  
pp. 117-123 ◽  
Author(s):  
Marina Guillén ◽  
Maria Dolors Benaiges ◽  
Francisco Valero
Keyword(s):  
Pichia Pastoris ◽  
Rhizopus Oryzae ◽  
Biochemical Properties ◽  
Recombinant Lipase

Optimization of the high-level production of Rhizopus oryzae lipase in Pichia pastoris

2001 ◽  
Vol 86 (1) ◽  
pp. 59-70 ◽  
Author(s):  
Stefan Minning ◽  
Alicia Serrano ◽  
Pau Ferrer ◽  
Carles Solá ◽  
Rolf D. Schmid ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Rhizopus Oryzae ◽  
Rhizopus Oryzae Lipase ◽  
High Level ◽  
Level Production

scholarly journals The Pichia pastoris PER6 gene product is a peroxisomal integral membrane protein essential for peroxisome biogenesis and has sequence similarity to the Zellweger syndrome protein PAF-1.

1996 ◽  
Vol 16 (5) ◽  
pp. 2527-2536 ◽  
Author(s):  
H R Waterham ◽  
Y de Vries ◽  
K A Russel ◽  
W Xie ◽  
M Veenhuis ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Membrane Protein ◽  
Sequence Similarity ◽  
Zellweger Syndrome ◽  
Methylotrophic Yeast ◽  
Integral Membrane Protein ◽  
Biochemical Properties ◽  
Podospora Anserina ◽  
Peroxisome Biogenesis ◽  
Membrane Spanning

We report the cloning of PER6, a gene essential for peroxisome biogenesis in the methylotrophic yeast Pichia pastoris. The PER6 sequence predicts that its product Per6p is a 52-kDa polypeptide with the cysteine-rich C3HC4 motif. Per6p has significant overall sequence similarity with the human peroxisome assembly factor PAF-1, a protein that is defective in certain patients suffering from the peroxisomal disorder Zellweger syndrome, and with car1, a protein required for peroxisome biogenesis and caryogamy in the filamentous fungus Podospora anserina. In addition, the C3HC4 motif and two of the three membrane-spanning segments predicted for Per6p align with the C3HC4 motifs and the two membrane-spanning segments predicted for PAF-1 and car1. Like PAF-1, Per6p is a peroxisomal integral membrane protein. In methanol- or oleic acid-induced cells of per6 mutants, morphologically recognizable peroxisomes are absent. Instead, peroxisomal remnants are observed. In addition, peroxisomal matrix proteins are synthesized but located in the cytosol. The similarities between Per6p and PAF-1 in amino acid sequence and biochemical properties, and between mutants defective in their respective genes, suggest that Per6p is the putative yeast homolog of PAF-1.

scholarly journals Truncated Prosequence of Rhizopus oryzae Lipase: Key Factor for Production Improvement and Biocatalyst Stability

Catalysts ◽  
2019 ◽  
Vol 9 (11) ◽  
pp. 961 ◽  
Author(s):  
Josu López-Fernández ◽  
Juan J. Barrero ◽  
M. Dolors Benaiges ◽  
Francisco Valero
Keyword(s):  
Amino Acids ◽  
Ionic Strength ◽  
Rhizopus Oryzae ◽  
Lipase Production ◽  
Biochemical Properties ◽  
Mature Sequence ◽  
Rhizopus Oryzae Lipase ◽  
Batch Cultures ◽  
Production Improvement ◽  
Key Factor

Recombinant Rhizopus oryzae lipase (mature sequence, rROL) was modified by adding to its N-terminal 28 additional amino acids from the C-terminal of the prosequence (proROL) to obtain a biocatalyst more suitable for the biodiesel industry. Both enzymes were expressed in Pichia pastoris and compared in terms of production bioprocess parameters, biochemical properties, and stability. Growth kinetics, production, and yields were better for proROL harboring strain than rROL one in batch cultures. When different fed-batch strategies were applied, lipase production and volumetric productivity of proROL-strain were always higher (5.4 and 4.4-fold, respectively) in the best case. rROL and proROL enzymatic activity was dependent on ionic strength and peaked in 200 mM Tris-HCl buffer. The optimum temperature and pH for rROL were influenced by ionic strength, but those for proROL were not. The presence of these amino acids altered lipase substrate specificity and increased proROL stability when different temperature, pH, and methanol/ethanol concentrations were employed. The 28 amino acids were found to be preferably removed by proteases, leading to the transformation of proROL into rROL. Nevertheless, the truncated prosequence enhanced Rhizopus oryzae lipase heterologous production and stability, making it more appropriate as industrial biocatalyst.

Evaluation of the performance of differently immobilized recombinant lipase B from Candida antarctica preparations for the synthesis of pharmacological derivatives in organic media

RSC Advances ◽  
2016 ◽  
Vol 6 (5) ◽  
pp. 4043-4052 ◽  
Author(s):  
Evelin A. Manoel ◽  
Julia M. Robert ◽  
Martina C. C. Pinto ◽  
Antonio C. O. Machado ◽  
Marina D. Besteti ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Candida Antarctica ◽  
Organic Media ◽  
Constitutive Promoter ◽  
Lipase B ◽  
Recombinant Lipase

This paper shows the production of lipase B fromCandida antarctica(LIPB) after cloning the gene that encoded it inPichia pastorisusing PGK as a constitutive promoter. The lipase was immobilized on different home-made supports for distinct reactions.

Production of a Rhizopus oryzae lipase from Pichia pastoris using alternative operational strategies

2007 ◽  
Vol 130 (3) ◽  
pp. 291-299 ◽  
Author(s):  
Anna Surribas ◽  
Rainer Stahn ◽  
José Luis Montesinos ◽  
Sven-Olof Enfors ◽  
Francisco Valero ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Rhizopus Oryzae ◽  
Rhizopus Oryzae Lipase ◽  
Operational Strategies
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