Raw starch saccharification

• US 4,618,579 A
• Filed: 09/28/1984
• Issued: 10/21/1986
• Est. Priority Date: 09/28/1984
• Status: Expired due to Fees

• Current Assignee: Genencor International Incorporated (International Flavors & Fragrances Incorporated)
• Original Assignee: Genencor International Incorporated (International Flavors & Fragrances Incorporated)
• Inventors: Pickens, Carl E., Niekamp, Carl W., Dwiggins, Bruce L.
• Primary Examiner(s): Shapiro, Lionel M.
• Application Number:
  US06/656,051
• Time in Patent Office: 753 Days
• Field of Search: 435/96, 435/99, 435/205
• US Class Current: 435/96
• CPC Class Codes: C12N 9/2428 Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase
• CPC Class Codes: C12P 19/20 produced by the action of an exo-1,4 alpha-glucosidase, e.g. dextrose
• CPC Class Codes: Y10S 435/911 using fungi

Abstract

A multi-step process is provided for solubilizing and saccharifying granular starch slurries of between 20 and 60% d.s. starch. In a first step, an enzyme mixture exhibiting raw starch hydrolyzing activity is added to the slurry and maintained in contact therewith until at least a substantial portion but less than all of the starch is solubilized. The resulting syrup is separated from the remaining insoluble fraction. In a second step, the insoluble fraction is used to prepare a second slurry containing less than about 20% d.s. starch, and this second slurry is solubilized with a raw starch hydrolyzing enzyme preparation until substantially all (e.g., at least about 95% or more) of the starch is solubilized. The pooled syrup from the two steps is over about 20 weight percent saccharides, at least about 95% of which is glucose.

Claims

1. A method of preparing a syrup containing, on a dry substance weight basis, a high percentage of glucose, said method comprising:

(a) preparing a first aqueous slurry of granular starch containing between about 20 and about 60 weight percent starch, d.s.;

(b) adjusting the pH and the temperature of said first slurry to values suitable to promote activity of a raw starch hydrolyzing enzyme preparation which is produced by a fungal organism and which catalyzes the hydrolysis of granular starch directly to glucose, said raw starch hydrolyzing enzyme preparation being characterized in that it catalyzes the hydrolysis of granular starch suspended in water at a concentration of about 15 percent by weight starch solids substantially completely to soluble glucose syrup solids containing at least about 97 percent by weight glucose, d.s., when the hydrolysis is carried out at a pH of about 5.0 to about 7.0 and a temperature of about 55°

C. and without added alpha amylase or added debranching enzyme of the pullulanase, isoamylase or beta amylase type, said enzyme preparation being further characterized in that it is separable by carboxymethyl cellulose into a first adsorbing fraction and a second non-adsorbing proteinaceous fraction, said first adsorbing fraction containing a glucoamylase enzyme (EC 3.2.1.3) that has an isoelectric point of about 8.0 or higher and said second non-adsorbing fraction having glucoamylase-protentiating activity that catalyzes the hydrolysis of granular starch;

(c) adding to said first slurry the raw starch hydrolyzing enzyme preparation in an amount sufficient to solubilize and saccharify a substantial portion of the starch in said first slurry;

(d) allowing said raw starch hydrolyzing enzyme preparation to solubilize and saccharify a substantial portion, but less than all, of the starch in said first slurry to yield a first syrup of at least about 22 weight percent saccharides, d.s.;

(e) separating said first syrup from the insolubles which remain and which contain unsolubilized starch from said first slurry;

(f) preparing a second aqueous slurry from said insolubles, said second slurry containing less than about 20 weight percent, d.s., starch;

(g) adjusting the pH and the temperature of said second slurry to values suitable to promote raw starch hydrolyzing enzyme activity;

(h) contacting said second slurry with the raw starch hydrolyzing enzyme preparation in an amount sufficient to solubilize and saccharify substantially all of the starch contained in said second slurry;

(i) allowing said raw starch hydrolyzing enzyme preparation to solubilize and saccharify substantially all of the starch in said second slurry to provide a second syrup; and

(j) separating said second syrup from insolubles contained therein.

2. A method according to claim 1 wherein the step (e) separation is made before the glucose content of the first syrup from step (d) drops below the 95 weight percent of the soluble saccharides in said first syrup.

3. A method according to claim 1 wherein step (d) is conducted over a period of about 48 hours or less.

4. A method according to claim 1 wherein said insolubles are removed from said syrups in steps (e) and (j) by centrifugation.

5. A method according to claim 1 wherein step (i) is carried out until at least about 95% of the starch in said second slurry is solubilized.

6. A method according to claim 1 wherein each of said first and second slurries contains between about 10 and about 200 ppm calcium.

7. A method according to claim 1 wherein said step (d) is conducted until at least about 25 percent of the starch in said first slurry is solubilized and saccharified.

8. The method according to claim 1 wherein said step (d) is conducted until at least about 50 percent of the starch in said first slurry is solubilized and saccharified.

9. The method according to claim 1 wherein at least about 60 percent of the starch in said first slurry is solubilized and saccharified in step (d) thereof.

10. The method according to claim 1 wherein said insolubles are removed from the syrups in steps (e) and (j) by centrifugation or filtration.

11. The method according to claim 1 wherein the first aqueous granular starch slurry contains from about 25 to about 40 weight percent granular starch on a total slurry weight basis.

12. The method according to claim 1 wherein the starch slurry of step (f) contains about 16 weight percent or less of granular starch on a total slurry weight basis.

13. The method of claim 1 wherein step (d) is terminated before the glucose content of the first syrup drops below about 97 weight percent on a soluble saccharide weight basis.

14. The method of claim 1 wherein said first and second syrups are pooled to provide a combined syrup containing in excess of 22 weight percent solubilized saccharide on a total syrup weight basis and in which at least about 95 weight percent of said saccharide is glucose.

15. The method of claim 1 wherein said first and second syrups are pooled to provide a combined syrup containing in excess of 24 weight percent solubilized saccharide on a total syrup weight basis, of which at least about 96 weight percent is glucose.

16. The method of claim 1 wherein the solubilization and saccharification of said first slurry is conducted at a pH of from about 5.0 to about 7.0 and at a temperature of from about 50°

to about 65°

C.

17. The method of claim 16 wherein the solubilization and saccharification of said second slurry is conducted at a pH of from about 5.0 to about 7.0 and at a temperature of from about 50°

to about 65°

C.

18. The method of claim 17 wherein the enzyme employed for the solubilization and saccharification of both the first and second starch slurries is a Humicola RSH enzyme.

19. The method of claim 1 wherein from about 2 to about 60 10 D.E. units of said RSH enzyme per gram of starch is employed to solubilize and saccharify each of said first and second aqueous starch slurries.

20. The method of claim 1 wherein the amount of RSH enzyme employed to solubilize and saccharify each of said first and second aqueous starch slurries is between about 5 and about 30 10 D.E. units.

21. The method of claim 1 wherein the granular starch employed is derived from corn.

22. A method of preparing a syrup containing, on a dry substance weight basis, a high percentage of glucose, said method comprising:

(a) preparing a first aqueous slurry of granular starch containing between about 20 and about 60 weight percent starch, d.s.;

(b) adjusting the pH and the temperature of said first slurry to values suitable to promote activity of a raw starch hydrolyzing enzyme preparation which is produced by a fungal organism that is a strain of the genus Humicola and which catalyzes the hydrolysis of granular starch directly to glucose, said raw starch hydrolyzing enzyme preparation being characterized in that it catalyzes the hydrolysis of granular starch suspended in water at a concentration of about 15 percent by weight starch solids substantially completely to soluble glucose syrup solids containing at least about 97 percent by weight glucose, d.s., when the hydrolysis is carried out at a pH of about 5.0 to about 7.0 and a temperature of about 55°

C. and without added alpha amylase or added debranching enzyme of the pullulanase, isoamylase or beta amylase type, said enzyme preparation being further characterized in that it is separable by carboxymethyl cellulose into a first adsorbing fraction and a second non-adsorbing proteinanceous fraction, said first adsorbing fraction containing a glucoamylase enzyme (EC 3.2.1.3) that has an isoelectric point of about 8.0 or higher and said second non-adsorbing fraction having glucoamylase-protentiating activity that catalyzes the hydrolysis of granular starch;

(c) adding to said first slurry the raw starch hydrolyzing enzyme preparation in an amount sufficient to solubilize and saccharify a substantial portion of the starch in said first slurry;

(d) allowing said raw starch hydrolyzing enzyme preparation to solubilize and saccharify a substantial portion, but less than all, of the starch in said first slurry to yield a first syrup of at least about 22 weight percent saccharides, d.s.;

(e) separating said first syrup from the insolubles which remain and which contain unsolubilized starch from said first slurry;

(f) preparing a second aqueous slurry from said insolubles, said second slurry containing less than about 20 weight percent, d.s., starch;

(g) adjusting the pH and the temperature of said second slurry to values suitable to promote raw starch hydrolyzing enzyme activity;

(h) contacting said second slurry with the raw starch hydrolyzing enzyme preparation in an amount sufficient to solubilize and saccharify substantially all of the starch contained in said second slurry;

(i) allowing said raw starch hydrolyzing enzyme preparation to solubilize and saccharify substantially all of the starch in said second slurry to provide a second syrup; and

(j) separating said second syrup from insolubles contained therein.

23. A method of preparing a syrup contaning, on a dry substance weight basis, a high percentage of glucose, said method comprising:

(a) preparing a first aqueous slurry of granular starch containing between about 20 and about 60 weight percent starch, d.s.;

(b) adjusting the pH and the temperature of said first slurry to values suitable to promote activity of a raw starch hydrolyzing enzyme preparation which is produced by a fungal organism that is a strain of the species Humicola grisea and which catalyzes the hydrolysis of granular starch directly to glucose, said raw starch hydrolyzing enzyme preparation being characterized in that it catalyzes the hydrolysis of granular starch suspended in water at a concentration of about 15 percent by weight starch solids substantially completely to soluble glucose syrup solids containing at least about 97 percent by weight glucose, d.s., when the hydrolysis is carried out at a pH of about 5.0 to about 7.0 and a temperature of about 55°

C. and without added alpha amylase or added debranching enzyme of the pullulanase, isoamylase or beta amylase type, said enzyme preparation being further characterized in that it is separable by carboxymethyl cellulose into a first adsorbing fraction and a second non-adsorbing proteinaceous fraction, said first adsorbing fraction containing a glucoamylase enzyme (EC 3.2.1.3) that has an isoelectric point of about 8.0 or higher and said second non-adsorbing fraction having glucoamylase-protentiating activity that catalyzes the hydrolysis of granular starch;

(c) adding to said first slurry the raw starch hydrolyzing enzyme preparation in an amount sufficient to solubilize and saccharify a substantial portion of the starch in said first slurry;

(d) allowing said raw starch hydrolyzing enzyme preparation to solubilize and saccharify a substantial portion, but less than all, of the starch in said first slurry to yield a first syrup of at least about 22 weight percent saccharides, d.s.;

(e) separating said first syrup from the insolubles which remain and which contain unsolubilized starch from said first slurry;

(f) preparing a second aqueous slurry from said insolubles, said second slurry containing less than about 20 weight percent, d.s., starch;

(g) adjusting the pH and the temperature of said second slurry to values suitable to promote raw starch hydrolyzing enzyme activity;

(h) contacting said second slurry with the raw starch hydrolyzing enzyme preparation in an amount sufficient to solubilize and saccharify substantially all of the starch contained in said second slurry;

(i) allowing said raw starch hydrolyzing enzyme preparation to solubilize and saccharify substantially all of the starch in said second slurry to provide a second syrup; and

(j) separating said syrup from insolubles contained therein.

24. A method of preparing a syrup containing, on a dry substance weight basis, a high percentage of glucose, said method comprising:

(a) preparing a first aqueous slurry of granular starch containing between about 20 and about 60 weight percent starch, d.s.;

(b) adjusting the pH and the temperature of said first slurry to values suitable to promote activity of a raw starch hydrolyzing enzyme preparation which is produced by a fungal organism that is a strain of the species Humicola grisea var. thermoidea and which catalyzes the hydrolysis of granular starch directly to glucose, said raw starch hydrolyzing enzyme preparation being characterized in that it catalyzes the hydrolysis of granular starch suspended in water at a concentration of about 15 percent by weight starch solids substantially completely to soluble glucose syrup solids containing at least about 97 percent by weight glucose, d.s., when the hydrolysis is carried out at a pH of about 5.0 to about 7.0 and a temperature to about 55°

C. and without added alpha amylase or added debranching enzyme of the pullulanase, isoamylase or beta amylase type, said enzyme preparation being further characterized in that it is separable by carboxymethyl cellulose into a first adsorbing fraction and a second non-adsorbing proteinaceous fraction, said first adsorbing fraction containing a glucoamylase enzyme (EC 3.2.1.3) that has an isoelectric point of about 8.0 or higher and said second non-adsorbing fraction having glucoamylase-protentiating activity that catalyzes the hydrolysis of granular starch;

(c) adding to said first slurry the raw starch hydrolyzing enzyme preparation in an amount sufficient to solubilize and saccharify a substantial portion of the starch in said first slurry;

(d) allowing said raw starch hydrolyzing enzyme preparation of solubilize and saccharify a substantial portion, but less than all, of the starch in said first slurry to yield a first syrup of at least about 22 weight percent saccharides, d.s.;

(e) separating said first syrup from the insolubles which remain and which contain unsolubilized starch from said first slurry;

(f) preparing a second aqueous slurry from said insolubles, said second slurry containing less than about 20 weight percent, d.s., starch;

(g) adjusting the pH and the temperature of said second slurry to values suitable to promote raw starch hydrolyzing enzyme activity;

(h) contacting said second slurry with the raw starch hydrolyzing enzyme preparation in an amount sufficient to solubilize and saccharify substantially all of the starch contained in said second slurry;

(i) allowing said raw starch hydrolyzing enzyme preparation to solubilize and saccharify substantially all of the starch in said second slurry to provide a second syrup; and

(j) separating said second syrup from insolubles contained therein.