Cell wall remnants and spheroplast regeneration in Mucor rouxii

Cabo, Amelia Pérez; Reyes, Emma; Novaes-Ledieu, Monique; Mendoza, Concepción García

doi:10.1016/s0007-1536(86)80055-9

Cited by 3 publications

(1 citation statement)

References 18 publications

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“…It is known that when obtaining fungal protoplasts, the formation of spheroplasts (cells with partial cell wall) may be observed and it often depends on the choice of lytic enzymes and duration time (Li et al 2017 ; Wang et al 1988 ). In turn, the presence of spheroplasts can increase the percentage of their regeneration (Wagner and Wilkinson 1993 ; Cabo et al 1986 ). Our goal was to obtain true intact protoplasts of G. zeae for their further mutagenesis, and for this purpose a lysing enzymes complex from T. harzianum contains β-glucanase, cellulase, protease, and chitinase activities necessary for complete removal of cell wall was chosen.…”

Section: Discussionmentioning

confidence: 99%

Ursodeoxycholic acid production by Gibberella zeae mutants

Kollerov

Donova

2022

AMB Expr

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Ursodeoxycholic acid (UDCA) is a highly demanded pharmaceutical steroid widely used in medicine. An ascomycete Gibberella zeae VKM F-2600 is capable of producing UDCA by 7β-hydroxylation of lithocholic acid (LCA). The present study is aimed at the improvement of the fungus productivity. The original procedures for the protoplast obtaining followed by UV mutagenesis and screening of ketoconazole-resistant mutant clones have been applied. The highest yield of G. zeae protoplasts was obtained when using the mycelium in the active growth phase, ammonium chloride as an osmotic stabilizer and treatment of the fungal cells by the lytic enzymes cocktail from Trichoderma hurzanium. The conditions for effective protoplast regeneration and the UV-mutagenesis were found to provide 6–12% survival rate of the protoplasts with superior number of possible mutations. Three of 27 ketoconazole-resistant mutant clones obtained have been selected due to their increased biocatalytic activity towards LCA. The mutant G. zeae M23 produced 26% more UDCA even at relatively high LCA concentration (4 g/L) as compared with parent fungal strain, and the conversion reached 88% (w/w). The yield of UDCA reached in this study prefers those ever reported. The results contribute to the knowledge on ascomycete mutagenesis, and are of importance for biotechnological production of value added cholic acids. Graphical Abstract

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