Method for preparation of fungal mutant with high hydrolytic activity

Patent metadata

Field	Value
Publication number	US-11261419-B2
Application number	US-201916719630-A
Country	US
Kind code	B2
Filing date	Dec 18, 2019
Priority date	Dec 19, 2018
Publication date	Mar 1, 2022
Grant date	Mar 1, 2022

Abstract

A method for preparing a hyper-cellulolytic catabolite derepressed mutants of ascomycetes fungus, especially variants of Penicillium funiculosum. Selection media used to isolate such variants include amorphous cellulose and a high concentration of glucose. Cellulase activities of mutant ID-10, in particular such as FPase and β-glucosidase were 1.5 times higher than Penicillium funiculosum MRJ-16 (parent). Furthermore, fungal mutant morphology was changed and no pH adjustment was required throughout the enzyme production process.

First claim

The invention claimed is:

1. A method of preparing a mutant fungal strain comprising:
(a) preparing a mutant fungal strain by contacting Penicillium funiculosum MRJ-16 with an aerobic culture medium followed by treatment with UV light or N-methyl-N′-nitro-Nnitrosoguanidine (NTG) or ethyl methanesulfonate (EMS) mutagens or in combination;
(b) screening for the mutant fungal strain having higher celluloytic activity as compared to the Penicillium funiculosum MRJ-16 by aerobic fermentation in an aerobic culture medium comprising amorphous cellulose and glucose at concentration of 1-4% (w/w) under conditions suitable for the production of enzymes;
(c) obtaining a mutant fungal strain Penicillium funiculosum -ID-10; and
(d) screening the efficiency of the enzyme produced by the mutant fungal strain Penicillium funiculosum -ID-10 by hydrolyzing biomass.

2. The method in of claim 1 , wherein the aerobic culture medium comprises 4% glucose.

3. The method in of claim 1 , wherein the fermentation in step (b) is carried in an aerated stirred tank having a glass jacketed vessel of 2 L and working volume of 1.8 L.

4. The method in of claim 1 , wherein the aerobic culture medium comprises ammonium sulphate 5 g/L, KH 2 PO 4 6 g/L, MgSO 4 *7H 2 O 1 g/L, CaCO 3 5 g/L, Glycerol 2.5 g/L, Corn steep solids 30 g/L, cellulose 30 g/L and Tween-80 2 ml/L.

5. The method of claim 1 , wherein the method is carried out in a fermenter, which was sterilized at 120° C. for 20 minutes and cooled at 30° C. along with maintaining pH of 5.5.

6. The method of claim 1 , wherein the enzyme produced in step (b) is a cellulase enzyme.

Assignees

• Indian Oil Corp Ltd

Inventors

• Adsul Mukund
• Sandhu Simranjeet Kaur
• Singhania Reeta Rani
• Mathur Anshu Shankar
• Gupta Ravi Prakash
• Tuli Deepak Kumar
• Puri Suresh Kumar

Classifications

• C12N2500/14

  Calcium; Ca chelators; Calcitonin · CPC title

• C12N1/22

  Processes using, or culture media containing, cellulose or hydrolysates thereof · CPC title

• C12N5/0018

  Culture media for cell or tissue culture (media for specific animal cell type C12N5/06) · CPC title

• C12N1/14 Primary

  Fungi (culture of mushrooms A01G18/00; as new plants A01H15/00); Culture media therefor · CPC title

• C12N2500/16

  Magnesium; Mg chelators · CPC title