DESC:AN INTEGRATED PROCESS FOR PRODUCTION OF CARBOHYDRATASES, ETHANOL, AND XYLITOL USING AN ISOLATED CANDIDA STRAIN ,CLAIMS:1. An isolated Candia tropicalis strain HP-CAT01 having accession number MTCC 25057.
2. A method of carbohydratases production using an isolated Candia strain HP-CAT01 having accession number MTCC 25057, said method comprising the steps of:
a. obtaining at least one lignocellulosic biomass;
b. pre-treating said lignocellulosic biomass to obtain a liquid hydrolysate, and a pre-treated solid biomass, wherein said pre-treating is with acid or alkali having v/v concentration of 0.2-5%, and said biomass w/v concentration is in the range of 10-20%;
c. separating said liquid hydrolysate, and said pre-treated solid biomass;
d. neutralizing said liquid hydrolysate by adjusting the pH in the range of 6-7; and
e. contacting Candia strain HP-CAT01 having accession number MTCC 25057 with minimal salts or at least one nitrogen source, 0.5-5% w/v pre-treated solid biomass, and said liquid hydrolysate of step (d) in a v/v percentage of 1-5% at a temperature in the range of 30-42°C for 24-96 hours under aerobic conditions to produce said carbohydratases.
3. The method as claimed in claim 2, wherein step (b) comprises contacting said lignocellulosic biomass with phosphoric or sulphuric acid at 145-155°C for 10-20 minutes, wherein the acid concentration is 0.5% and solid loading w/v concentration is 10%.
4. The method as claimed in claim 2, wherein said carbohydratases are selected from the group consisting of endoglucanase, exoglucanase, beta-glucosidase, xylanase, and combinations thereof.
5. A method of production of at least one sugar using an isolated Candia strain HP-CAT01 having accession number MTCC 25057, said method comprising the steps of:
a. obtaining at least one lignocellulosic biomass;
b. pre-treating said lignocellulosic biomass to obtain a liquid hydrolysate, and a pre-treated solid biomass, wherein said pre-treating is with acid or alkali having v/v concentration of 0.2-5%, and said biomass w/v concentration is in the range of 10-20%;
c. separating said liquid hydrolysate, and said pre-treated solid biomass;
d. neutralizing said liquid hydrolysate by adjusting the pH in the range of 6-7;
e. contacting Candia strain HP-CAT01 having accession number MTCC 25057 with minimal salts or at least one nitrogen source, 0.5-5% w/v pre-treated solid biomass, and said liquid hydrolysate of step (d) in a v/v percentage of 1-5% at a temperature in the range of 30-42° for 24-96 hours under aerobic conditions to produce said carbohydratases;
f. enriching said carbohydratases to obtain concentrated carbohydratases; and
g. incubating said concentrated carbohydratases with said pre-treated solid biomass at a temperature in the range of 48-52°C for 24-96 hours under aerobic conditions to produce said sugar.
6. The method as claimed in claim 5, wherein said sugar is 70-80% hexose.
7. The method as claimed in claim 5, wherein said carbohydratases are selected from the group consisting of endoglucanase, exoglucanase, beta-glucosidase, xylanase, and combinations thereof.
8. The method as claimed in claim 5, wherein step (b) comprises contacting said lignocellulosic biomass with phosphoric or sulphuric acid at 145-155°C for 10-20 minutes, wherein the acid concentration is 0.5% and solid loading w/v concentration is 10%.
9. A method of ethanol production using an isolated Candia strain HP-CAT01 having accession number MTCC 25057, said method comprising the steps of:
a. obtaining at least one lignocellulosic biomass;
b. pre-treating said lignocellulosic biomass to obtain a liquid hydrolysate, and a pre-treated solid biomass, wherein said pre-treating is with acid or alkali having v/v concentration of 0.2-5%, and said biomass w/v concentration is in the range of 10-20%;
c. separating said liquid hydrolysate, and said pre-treated solid biomass;
d. neutralizing said liquid hydrolysate by adjusting the pH in the range of 6-7;
e. contacting Candia strain HP-CAT01 having accession number MTCC 25057 with minimal salts or at least one nitrogen source, 0.5-5% pre-treated solid biomass, and said liquid hydrolysate of step (d) in a v/v percentage of 1-5% at a temperature in the range of 30-42° for 24-96 hours under aerobic conditions to produce said carbohydratases;
f. enriching said carbohydratases to obtain concentrated carbohydratases;
g. incubating said concentrated carbohydratases with said pre-treated solid biomass at a temperature in the range of 48-52°C for 24-96 hours under aerobic conditions to produce said sugar; and
h. contacting said Candida strain with said sugar at 30-42°C for 24 -48 hours under aerobic or anaerobic conditions to produce ethanol.
10. The method as claimed in claim 9, wherein said sugar is predominantly hexose.
11. The method as claimed in claim 9, wherein said carbohydratases are selected from the group consisting of endoglucanase, exoglucanase, beta-glucosidase, xylanase, and combinations thereof.
12. The method as claimed in claim 9, wherein step (b) comprises contacting said lignocellulosic biomass with phosphoric or sulphuric acid at 145-155°C for 10-20 minutes, wherein the acid concentration is 0.5% and solid loading w/v concentration is 10%.
13. A method of xylitol production using an isolated Candida strain HP-CAT01 having accession number MTCC 25057, said method comprising the steps of:
a. obtaining at least one lignocellulosic biomass;
b. pre-treating said lignocellulosic biomass to obtain a liquid hydrolysate, and a pre-treated solid biomass, wherein said pre-treating is with acid or alkali having v/v concentration of 0.2-5%, and said biomass w/v concentration is in the range of 10-20%;
c. separating said liquid hydrolysate, and said pre-treated solid biomass;
d. neutralizing said liquid hydrolysate by adjusting the pH in the range of 6-7; and
e. contacting said isolated Candida strain HP-CAT01 having accession number MTCC 25057 with minimal salts or at least one nitrogen source, and said liquid hydrolysate at a temperature in the range of 30-42°C for 24-72 hours under aerobic conditions to promote xylitol production.
14. The method as claimed in claim 13, wherein step (b) comprises contacting said lignocellulosic biomass with phosphoric or sulphuric acid at 145-155°C for 10-20 minutes, wherein the acid concentration is 0.5% and solid loading w/v concentration is 10%.
15. An integrated method of ethanol and xylitol production using an isolated Candida strain HP-CAT01 having accession number MTCC 25057, said method comprising the steps of:
a. obtaining at least one lignocellulosic biomass;
b. pre-treating said lignocellulosic biomass to obtain a liquid hydrolysate, and a pre-treated solid biomass, wherein said pre-treating is with acid or alkali having v/v concentration of 0.2-5%, and said biomass w/v concentration is in the range of 10-20%;
c. separating said liquid hydrolysate, and said pre-treated solid biomass;
d. neutralizing said liquid hydrolysate by adjusting the pH in the range of 6-7;
e. contacting Candia strain HP-CAT01 having accession number MTCC 25057 with minimal salts or at last one nitrogen source 0.5-5% pre-treated solid biomass, and said liquid hydrolysate of step (d) in a v/v percentage of 1-5% at a temperature in the range of 30-42° for 24-96 hours under aerobic conditions to produce said carbohydratases;
f. separating said Candida strain population from (e) to obtain a cell free liquid hydrolysate;
g. contacting said pre-treated solid biomass to said cell free liquid hydrolysate at a temperature in the range of 45-55°C for 24-96 hours under aerobic conditions to promote production of sugars; and
h. contacting said isolated Candida strain HP-CAT01 having accession number MTCC 25057 with sugars from step (g) at a temperature in the range of 30-37°C for 24–48 hours in aerobic conditions to promote ethanol and xylitol production.
16. The method as claimed in claim 15, further comprising the step of fractional distillation to separate ethanol, and xylitol.
17. The method as claimed in claim 15, wherein said carbohydratases are selected from the group consisting of endoglucanase, exoglucanase, beta-glucosidase, xylanase, and combinations thereof.
18. The method as claimed in claim 15, wherein step (b) comprises contacting said lignocellulosic biomass with phosphoric or sulphuric acid at 145-155°C for 10-20 minutes, wherein the acid concentration is 0.5% and solid loading w/v concentration is 10%.
19. The method as claimed in any of the claims 2-18, wherein said minimal salts are selected from the group consisting of 0.5 g/L MgSO4.7H2O, 0.5 g/: KCl, 1 g/L KH2PO4, 2.5 g/L NaNO3, and combinations thereof.
20. The method as claimed in any of the claims 2-18, wherein said at least one nitrogen source concentration is in the range of 0.5-2%.
21. The method as claimed in any of the claims 2-18, wherein said isolated Candia tropicalis strain HP-CAT01 having accession number MTCC 25057 is active in a pH range of 4-10, a temperature range of 30–42 0C, and inhibitory compounds concentration range of 0.1–3 g/L.
22. The method as claimed in any of the claims 2-18, wherein said lignocellulosic biomass is selected from the group consisting of wheat straw, rice straw, sorghum straw, agricultural feedstock, carboxymethylcellulose (CMC), and combinations thereof.
23. An isolated Candida tropicalis strain HP-CAT01 having accession number MTCC 25057 for use in production of ethanol from lignocellulosic biomass.
24. An isolated Candida tropicalis strain HP-CAT01 having accession number MTCC 25057 for use in production of xylitol from lignocellulosic biomass.
25. An isolated Candida tropicalis strain HP-CAT01 having accession number MTCC 25057 for use in production of xylitol and ethanol from lignocellulosic biomass.
26. An isolated Candida tropicalis strain HP-CAT01 having accession number MTCC 25057 for use in production of carbohydratases from lignocellulosic biomass.
27. An isolated Candida tropicalis strain HP-CAT01 having accession number MTCC 25057 for use in production of sugars from lignocellulosic biomass.
28. A bio-plant for production of bioethanol, xylitol, value added products, sugars, and enzymes from lignocellulosic biomass using an isolated Candia tropicalis strain HP-CAT01 having accession number MTCC 25057.
Dated this 2nd September 2015