Bio-refinery waste utilization for enzyme production using novel penicillium funiculosum MRJ-16 fungal strain

Patent metadata

Field	Value
Publication number	US-11242549-B2
Application number	US-202016785599-A
Country	US
Kind code	B2
Filing date	Feb 8, 2020
Priority date	Apr 17, 2019
Publication date	Feb 8, 2022
Grant date	Feb 8, 2022

Abstract

The present invention relates to a method for obtaining a high titer of enzyme mixture comprising cellulases, hemicellulases and β-glucosidases in reutilization of waste water generated during hot water extraction of lignocellulosic biomass or biorefinery waste water using Penicillium funiculosum MRJ-16 mutant strain. The cellulose or lignocellulosic biomass used in the fermentation process is selected from the group consisting of rice straw, wheat straw, corn stover, cotton stalk or a combination thereof. The enzyme mixture obtained by the present process is used for the saccharification of acid pretreated lignocellulosic biomass.

First claim

We claim:

1. A method for obtaining an enzyme mixture comprising cellulases, hemicellulases and beta-glucosidases in re-utilization of waste water, the method comprising the steps of:
(a) preparing fermentation media comprising 3-5 g/L ammonium sulfate, 4-6 g/L KH 2 PO 4 , 0.5-1 g/L MgSO 4 -7H 2 O, 2.5-5 g/L CaCO 3 , 2-3 g/L glycerol, 25-35 g/L corn steep solids, 25-40 g/L carbon source, 1-2 ml/L polysorbate 80, and waste water obtained during pre-treatment of lignocellulosic biomass or biorefinery waste water;
(b) inoculating the media of step (a) with a liquid seed culture of Penicillium funiculosum MRJ-16 of Microbial Type Culture Collection (MTCC) Accession Number 25142;
(c) fermenting the Penicillium funiculosum MRJ-16 culture of step (b) in an aerated fermenter; and
(d) harvesting an enzyme mixture after centrifuging the fermented culture broth of step (c).

2. The method as claimed in claim 1 , wherein the waste water comprises waxes, proteins, phenolics and inhibitors.

3. The method as claimed in claim 2 , wherein the inhibitors in the waste water are organic acid, phenol, hydroxyl methyl furfural (HMF), formic acid, acetic acid and furfural.

4. The method as claimed in claim 1 , wherein the carbon source in the fermentation media of step (a) is selected from the group consisting of cellulose, acid pretreated lignocellulosic biomass, and a combination thereof.

5. The method as claimed in claim 1 , wherein the fermentation process is carried out at a temperature in the range of 25-35° C., pH in the range of 4-6, aeration in the range of 0.5-2.5 WM, and dissolved oxygen above 30%.

6. The method as claimed in claim 1 , wherein the fermentation is carried out for a time period of 80-110 hours.

7. The method as claimed in claim 4 , wherein the cellulose or lignocellulosic biomass is selected from the group consisting of rice straw, wheat straw, corn stover, cotton stalk, and a combination thereof.

8. The method as claimed in claim 1 , wherein the enzyme mixture obtained is used for the saccharification of acid pretreated lignocellulosic biomass.

Assignees

• Indian Oil Corp Ltd
• Dept Of Biotechnology India

Inventors

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

Classifications

• C12P19/14 Primary

  produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase} · CPC title

• C12P21/02 Primary

  having a known sequence of two or more amino acids, e.g. glutathione · CPC title

• C12N1/145

  Fungi isolates · CPC title

• C12P19/16

  produced by the action of an alpha-1, 6-glucosidase, e.g. amylose, debranched amylopectin (non-biological hydrolysis of starch C08B30/00) · CPC title

• C12P2201/00

  Pretreatment of cellulosic or lignocellulosic material for subsequent enzymatic treatment or hydrolysis · CPC title