Starch processing is a well-established area for the commercial application of enzymes.  For instance, high fructose corn syrup (HFCS) is made using a range of enzymes. HFCS act as a substitute for ordinary sugar in food and beverage processing, and are used extensively in, for example, the soft drinks industry.  For more information on HFCS and many other valuable products derived from starch by enzymatic processes, please connect to the starch website.

Novozymes has invested heavily into enzyme research in the Starch field, and performed intensive development work on application processes. Naturally, numerous inventions have been made throughout this process, and Novozymes has obtained a significant patent portfolio.  The inventions claimed in the patents include new enzymes isolated from natural sources, enzymes improved by protein engineering, immobilization of enzymes and processes for commercial application of enzymes.  The enzymes are the typical enzymes used for production of HFCS, such as amylases, glucoamylases, pullulanases and glucose isomerases, as well as other starch-related enzymes, including cyclodextrin glucosyl transferases. 

We are convinced that many of the enzymes and processes, which we claim in our ?starch? patents, also will have commercial potential outside the traditional range of enzymatically derived starch products.  Perhaps, you have the vision and knowledge to pursue such new applications, based on Novozymes proprietary technologies.
 
A selected number of the patent titles applicable to the starch industry are shown below.  If you are interested in one of these patents, or in a Novozymes starch patent not shown here, we welcome you to contact us to discuss a possible license to your company.

• Immobilization of Biologically Active Material with Glutaraldehyde and Polyazaetidine

• Cyclomaltodextrin glucanotransferase variants

• Novel cyclomaltodextrin glucanotransferase (CGTase) variants
  
  

Title of Technology:  Immobilization of Biologically Active Material with Glutaralde-hyde and Polyazaetidine

Abstract: Immobilized biologically active material in particle form is pre-pared by cross-linking with glutaraldehyde and polyazetidine.

An aqueous dispersion or solution of biologically active mate-rial is partially cross-linked with glutaraldehyde, a wet pasty mass is recovered by dewatering and the mas is sub-divided into discrete particles. 

A polyazetidine prepolymer is added before, at the beginning or subsequent to partially cross-linking but prior to subdividing the pasty mass into particles, and the prepolymer is allowed to cross-link.

Detailed Description:  In brief, the method of this technology comprises partially cross-linking enzymatically active microorganism cells or some other biological material in aqueous solution or suspen-sion through reaction with glutaraldehyde, followed by dewa-tering of the resulting flocculated solids, resulting in a pasty consistency mass of a (consistency) and coherency suitable for particle shaping. Then, (wet) particles of desired shape and size are generated from the mixture followed by drying.

The polyazetidine prepolymer solution is incorporated before or after dewatering, more preferably the former. · Curing of the polyazetidine prepolymer which occurs during the drying step, converts the pasty mass particles into cured particles of high physical strength.

Immobilized enzyme products prepared according to the in-vention e mployed in packed bed exhibits a very small pres-sure drop (e.g., a pressure drop which is only around 50%of a comparable prior art product) and a high physical strength and resistance against abrasion, and moreover, the im-moblized enzyme products are relatively inexpensive to make due to the high yield of usable particles.

Also, it has been found that preferred embodiment immobi-lized enzyme product prepared according to the invention ex-hibit a high volumetric activity.

Patent References

Patent Number:	Title of Patent:	Year of Issue:
EP297912B1	Process for immobilization	1989
US-Re.33441	Immobilization of biologically acive material with glutaraldehyde and polyazetidine	1990
JP2641934B2		1997
KR9700186B1		1997

 

 

 

 

 

   
Title of Technology:  · Cyclomaltodextrin glucanotransferase variants·

 
Abstract: Th e present technology relates to variants of cyclomaltodextrin glucanotransferase (CGTase). More specifically the technology relates to a method of modifying the substrate binding and/or product selectivity of a precursor CGTase enzyme, and CGTase variants derived from a precursor CGTase enzyme by substitution, insertion and/or deletion of one or more amino acid residue(s), which amino acid residue(s) holds a position close to the substrate.

Moreover, the technology relates to DNA constructs encoding the CGTase variants, expression vectors, host cells and meth-ods of producing the CGTase variants.

Detailed Description:  Cyclomaltodextrin glucanotransferase (E.C. 2.4.1.19), also des-ignated cyclodextrin glucanotransferase or cyclodextrin glyco-syltransferase (CGTase), catalyses the conversion of starch and similar substrates into cyclomaltodextrins via an in-tramolecular transglycosylation reaction, thereby forming cyc-lomaltodextrins, in the following termed cyclodextrins, of various sizes.

Commercially most important are cyclodextrins of 6, 7 and 8 glucose units, which are termed a-, b- and g-cyclodextrins, re-spectively. Commercially less important are cyclodextrins of 9, 10, and 11 glucose units, which are termed d-, e-, and z-cyclodextrins, respectively.· Cyclodextrins are thus cyclic glucose oligomers with a hydro-phobic internal cavity. They are able to form inclusion com-plexes with many small hydrophobic molecules in aqueous so-lutions, resulting in changes in physical properties, e.g. in-creased solubility and stability and decreased chemical reactiv-ity and volatility. Cyclodextrins find applications particularly in the food, cosmetic, chemical and pharmaceutical industries.

This technology provides novel variants of CGTases, which variants, when compared to the precursor enzyme, show in-creased product selectivity and/or reduced product inhibition.

Accordingly, in its first aspect, the technology provides a method of modifying the substrate binding and/or product selec-tivity of a precursor CGTase enzyme, which method comprises substitution, insertion and/or deletion of one or more amino acid residue(s) of the precursor enzyme, which amino acid resi-due(s) holds a position close to the substrate.

In another aspect, the technology provides a CGTase variant derived from a precursor CGTase enzyme by substitution, in-sertion and/or deletion of one or more amino acid residue(s), which amino acid residue(s) holds a position close to the sub-strate.

The technology further provides for DNA constructs, recombi-nant expression vectors, host cells, a method of producing the CGTase variant, and for the CGTase variants? use in processes for the manufacture of cyclodextrins and linear oligosaccha-rides.

Patent References

Patent Number:	Title of Patent:	Year of Issue:
WO9633267	Cyclomaltodextrin glucanotransferase vari-ants
US6004790	Cyclomaltodextrin glucanotransferase vari-ants	1999

 

 

 

 

Title of Technology:  Novel cyclomaltodextrin glucanotransferase (CGTase) vari-ants

Abstract:& nbsp;· The present technology provides novel CGTase variants of increased product specificity in which one or more of the amino acid residues corresponding to the following positions have been introduced by substitution and/or insertion (CGTase Num-bering):

(i)  Position 47Cys/Asp/Glu/Phe/Gly/Ile/Lys/Asn/Pro/Arg/Ser/Thr/Val/Trp/Tyr;

(ii)  Position 145Asp/His/Ile/Asn/Gln/Val;

(iii)  Position 146His/Lys/Leu/Thr/Val/Tyr;(iv)  Position 147Cys/Asp/Glu/Asn/Gln; (v)  Position 196Cys/Glu/Phe/Gly/His/Ile/Lys/Leu/Met/Pro/Gln/Arg/Thr/Val/Trp/Tyr and/or

(vi)  Position 371Cys/Glu/Phe/His/Ile/Lys/Leu/Met/Gln/Arg/Thr/Val/Trp.

Detailed Description:  Cyclomaltodextrin glucanotransferase (E.C. 2.4.1.19), also des-ignated cyclodextrin glucanotransferase or cyclodextrin glyco-syltransferase (CGTase), catalyses the conversion of starch and similar substrates into cyclomaltodextrins via an in-tramolecular transglycosylation reaction, thereby forming cyc-lomaltodextrins, in the following termed cyclodextrins, of various sizes.

Commercially most important are cyclodextrins of 6, 7 and 8 glucose units, which are termed a-, b- and g-cyclodextrins, re-spectively. Commercially less important are cyclodextrins of 9, 10, and 11 glucose units, which are termed d-, e-, and z-cyclodextrins, respectively.

Cyclodextrins are thus cyclic glucose oligomers with a hydro-phobic internal cavity. They are able to form inclusion com-plexes with many small hydrophobic molecules in aqueous so-lutions, resulting in changes in physical properties, e.g. in-creased solubility and stability and decreased chemical reactiv-ity and volatility. Cyclodextrins find applications particularly in the food, cosmetic, chemical and pharmaceutical industries.

CGTase variants showing an inc reased product specificity with respect to the production of a-cyclodextrin are provided, in which variants one or more of the amino acid residues corre-sponding to the following positions have, preferably, been intro-duced by substitution and/or insertion (CGTase Numbering):

(i)  Position 47Phe/Lys/Arg/Trp/Tyr;

(ii)  Position 145Asp/His/Asn/Gln;

(iii)  Position 146His/Lys/Leu/Thr/Val/Tyr;

(iv)  Position 147Cys/Asp/Glu/Asn/Gln;

(v)  Position 196Cys/Glu/Phe/Gly/His/Ile/Lys/Leu/Met/Pro/Gln/Arg/Thr/Val/Trp/Tyr/ and/or

(vi)  Position 371Cys/His/Lys/Arg/Thr.

CGTase variants showing an increased product specificity with respect to the production of b-cyclodextrin are provided, in which variants one or more of the amino acid residues corre-sponding to the following positions have, preferably, been intro-duced by substitution and/or insertion (CGTase Numbering):

(i)  Position 47Cys/Asp/Glu/Phe/Gly/Ile/Asn/Pro/Ser/Thr/Val/Trp/Tyr;

(ii)  Position 145Asp/Ile/Asn/Val;

(iii)  Position 147Glu;

(iv)  Position 196Cys/Glu/Phe/Gly/His/Ile/Lys/Leu/Met/Pro/Gln/Arg/Thr/Val/Trp/Tyr and/or

(v)  Position 371Cys/Glu/Phe/His/Ile/Lys/Leu/Met/Gln/Arg/Thr/Val/Trp.

In yet another preferred embodiment, CGTase variants show-ing an increased product specificity with respect to the pro-duction of g-cyclodextrin are provided, in which variants one or more of the amino acid residues corresponding to the following positions have, preferably, been introduced by substitution and/or insertion (CGTase Numbering):

(i)  Position 47Cys/Asp/Glu/Phe/Gly/Ile/Asn/Pro/Ser/Thr/Val/Trp/Tyr;

(ii)  Position 145Asp/Ile/Asn/Val;

(iii)  Position 147Glu;

(iv)  Position 196Cys/Glu/Phe/Gly/His/Ile/Lys/Leu/Met/Pro/Gln/Arg/Thr/Val/Trp/Tyr and/or

(v)  Position 371Cys/Glu/Phe/His/Lys/Met/Gln/Arg/Thr/Trp.

 

Patent References

Patent Number:	Title of Patent:
WO9915633	Novel cyclomaltodextrin glucanotransferase variants