High-level constitutive expression in Pichia pastoris and one-step purification of phospholipase D from cowpea (Vigna unguiculata L. Walp)

Cowpea (Vigna unguiculata) is a neglected crop native to Africa, with an outstanding potential to contribute to the major challenges in food and nutrition security, as well as in agricultural sustainability. Two major issues regarding cowpea research have been highlighted in recent years—the establishment of core collections and the characterization of landraces—as crucial to the implementation of environmentally resilient and nutrition-sensitive production systems. In this work, we have collected, mapped, and characterized the morphological attributes of 61 cowpea genotypes, from 10 landraces spanning across six agro-ecological zones and three provinces in Mozambique. Our results reveal that local landraces retain a high level of morphological diversity without a specific geographical pattern, suggesting the existence of gene flow. Nevertheless, accessions from one landrace, i.e., Maringué, seem to be the most promising in terms of yield and nutrition-related parameters, and could therefore be integrated into the ongoing conservation and breeding efforts in the region towards the production of elite varieties of cowpea.

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

Journal of Biotechnology ◽

10.1016/s0168-1656(00)00402-8 ◽

2001 ◽

Vol 86 (1) ◽

pp. 59-70 ◽

Cited By ~ 124

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

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Mutations Causing Constitutive Invertase Synthesis in Yeast: Genetic Interactions with snf Mutations

Genetics ◽

10.1093/genetics/115.2.247 ◽

1987 ◽

Vol 115 (2) ◽

pp. 247-253

Author(s):

Lenore Neigeborn ◽

Marian Carlson

Keyword(s):

Regulatory Region ◽

Constitutive Expression ◽

Genetic Interactions ◽

The Third ◽

Recessive Mutations ◽

Constitutive Mutation ◽

Complementation Groups ◽

High Level ◽

Yeast Genetic ◽

Very High

ABSTRACT We have selected 210 mutants able to grow on sucrose in the presence of 2-deoxyglucose. We identified recessive mutations in three major complementation groups that cause constitutive (glucose-insensitive) secreted invertase synthesis. Two groups comprise alleles of the previously identified HXK2 and REG1 genes, and the third group was designated cid1 (constitutive invertase derepression). The effect of cid1 on SUC2 expression is mediated by the SUC2 upstream regulatory region, as judged by the constitutive expression of a SUC2-LEU2-lacZ fusion in which the LEU2 promoter is under control of SUC2 upstream sequences. A cid1 mutation also causes glucose-insensitive expression of maltase. The previously isolated constitutive mutation ssn6 is epistatic to cid1, reg1 and hxk2 for very high level constitutive invertase expression. Mutations in SNF genes that prevent derepression of invertase are epistatic to cid1, reg1 and hxk2; we have previously shown that ssn6 has different epistasis relationships with snf mutations. The constitutive mutation tup1 was found to resemble ssn6 in its genetic interactions with snf mutations. These findings suggest that CID1, REG1 and HXK2 are functionally distinct from SSN6 and TUP1.

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