Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase
Patent
1992-11-25
1999-01-26
Wax, Robert A.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Hydrolase
435 691, 435 72, 4352523, 43525231, 4351723, 43525411, 43525421, 4352543, 4352546, 4353201, 536 232, 935 14, 935 28, 935 68, 935 69, C12N 924, C12N 1556, C12N 1574, C12N 1580
Patent
active
058637834
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to the field of molecular biology. In particular, the present invention relates to the cloning and expression of genes encoding enzymes of fungal origin.
BACKGROUND OF THE INVENTION
The composition of the plant cell wall is complex and variable. Polysaccharides are mainly found in the form of long chains of cellulose (the main structural component of the plant cell wall), hemicellulose (comprising various .beta.-xylan chains) and pectic substances (consisting of galacturonans and rhamnogalacturonans; arabinans; and galactans and arabinogalactans). From the standpoint of the food industry, the pectic substances, arabinans in particular, have become one of the most important constituents of plant cell walls (Whitaker, J. R. (1984) Enzyme Microb. Technol., 6, 341).
Arabinans consist of a main chain of .alpha.-L-arabinose subunits linked .alpha.-(1.fwdarw.5) to one another. Side chains are linked .alpha.-(1.fwdarw.3) or sometimes .alpha.-(1.fwdarw.2) to the main .alpha.-(1.fwdarw.5)-L-arabinan backbone. In apple, for example, one third of the total arabinose is present in the side chains. The molecular weight of arabinan is normally about 15 kDa.
Enzymes capable of degrading arabinans are becoming increasingly important to the food industry. In juice production, for example, the demand to increase yields in order to reduce production costs has necessitated the modification of traditional processes. The utilization of enzymatic pre-treatments of the fruit pulp before pressing with specific enzymatic products drastically improves the juice yield by solubilizing the cell wall polysaccharides.
However, a persistent turbidity commonly referred to as "arabinan haze" has been a source of problems in the production of concentrated juices. The arabinan haze is more often present in concentrated juice than in non-concentrated juice. This may indicate that water activity has an influence on the solubility of arabinan. Furthermore, it has been found that this haze is soluble between 60.degree. and 80.degree. C.
It has also been found that while branched arabinan is soluble in concentrated chilled apple and pear juices, the linear, debranched .alpha.-(1.fwdarw.5)-L-arabinan is much less soluble. This debranched .alpha.-(1.fwdarw.5)-L-arabinan is formed from the L-arabinan by the action of an arabinan-degrading enzyme present in the commercial pectic enzyme preparations from Aspergillus niger, commonly used to increase juice yield after pulp treatment but before pressing.
On the other hand, debranched arabinans are considered desirable for certain other applications. WO 90/06343 discloses the debranching of sugar beet araban by the action of an .alpha.-L-arabinofuranosidase, free of endo arabinanase activity, which is isolated from a culture filtrate of Aspergillus niger or from a commercial pectinase mixture using ion-exchange and gel filtration chromatography procedures. The debranched araban may be used as a fat substitute in foods.
Arabinan-degrading enzymes are known to be produced by a variety of plants and microorganisms, among these, fungi such as those of the genera Aspergillus, Corticium, Rhodotorula (Kaji, A. (1984) Adv. Carbohydr. Chem. Biochem., 42, 383), Dichotomitus (Brillouet et al. (1985) Carbohydrate Research, 144, 113), Ascomycetes and Basidomycetes (Sydow, G. (1977) DDR Patent Application No. 124,812).
In particular, the filamentous fungus Aspergillus niger is known to produce three different arabinan-degrading enzymes: an .alpha.-L-arabinanase having a molecular weight of approximately 35 kDa and two .alpha.-L-arabinofuranosidases having molecular weights of approximately 118 and 60 kDa, respectively, (Rombouts et al. (1988) Carbohydrate 157, 23) reports molecular weights of 43, 83 and 67 kDa for these same three enzymes, respectively.!
The 35 kDa arabinanase (also known as ABN A) has endo activity and exclusively cleaves 1.fwdarw.5 linkages. The activity of this enzyme decreases as the 1,5-.alpha.-L-arabinan sequences become shorter and the concentration of arabinose di
REFERENCES:
patent: 4703008 (1987-10-01), Lin
patent: 4757006 (1988-07-01), Toole, Jr. et al.
patent: 4760025 (1988-07-01), Estell et al.
patent: 5013652 (1991-05-01), Strausberg et al.
patent: 5075227 (1991-12-01), Hagen
patent: 5108918 (1992-04-01), Groenen et al.
patent: 5143844 (1992-09-01), Abrahmsen et al.
patent: 5188958 (1993-02-01), Moloney et al.
patent: 5246849 (1993-09-01), Bryan et al.
patent: 5324653 (1994-06-01), Van Eekelen et al.
patent: 5336611 (1994-08-01), Van Eekelen et al.
patent: 5340735 (1994-08-01), Christianson et al.
patent: 5340738 (1994-08-01), Lambeir et al.
patent: 5384257 (1995-01-01), Lambeir et al.
patent: 5397705 (1995-03-01), Zukowski et al.
patent: 5482849 (1996-01-01), Branner et al.
Whitaker et al., "Pectic substances, peptic enzymes and haze formation in fruit juices" Enzyme Microb. Technol. (1984)6:341-349.
Kaji, A., "L-Arabinosidases" Adv. Carbohydrate Chem. Biochem. (1984) 42:383-394.
Brillouet et al., "Production, purification, and properties of an .alpha.-L-arabinofuranosidase from Dichomitus squalens" Carbohydrate Res. (1985) 144:113-126.
Rombouts et al., "The arabinanases of Aspergillus niger-purification and characterisation of two .alpha.-L-arabinofuranosidases and an endo-1,5-.alpha.-L-arabinanase" Carbohydrate Polymers (1988) 9:25-47.
van der Veen et al., "Induction, purification and characterisation of arabinases produced by Aspergillus niger" Arch. Microbiol. (1991) 157:23-28.
Gunata et al., "Purification and some properties of an .alpha.-L-arabinofuranosidase from Aspergillus niger. Action on grape monoterpenyl arabinofuranosylglucosides" J. Agric. Food Chem. (1990) 38:772-776.
Schwarz et al., "Xylan degradation by the thermophile clostridium stercorarium: cloning and expression of xylanase, .beta.-D-xylosidase, and .alpha.L-arabinofuranosidase genes in Escherichia coli" Biochem. Biophys. Res. Commun. (1990) 170(1):368-374.
Whitehead et al., "The genes for three xylan-degrading activities from Bacteriodes ovatus are clustered in a 3.8-kilobase region" J. Bacteriol. (1990) 172(5):2408-24112.
Karimi et al., "Comparative study of some microbial arabinan-degrading enzymes" J. Indust. Microbiol. (1989) 4(3):173-180.
Lee et al., "Purification and characterization of an .alpha.-L-arabinofuranosidase from Clostriduim acetobutylicum ATCC 824" Can. J. Microbiol. (1987) 33:1011-1016.
Jaye, M., et al., Nucleic Acids Research, vol. 11, No. 8, "Isolation of a human anti-haemophiliac factor IX cDNA clone using a unique 52-base synthetic oligonucleotide probe deduced from the amino acid sequence of bovine factor IX", pp. 2325-2335, 1983.
Ullrich. A., et al., The EMBO Journal, vol. 3, No. 2, "Isolation of the human insulin-like growth factor I gene using a single synthetic DNA probe", pp. 361-364, 1984.
Lehtovaara, P. M., et al., Protein Engineering, vol. 2, "A new method for random mutagenesis of complete genes: enzymatic generation of mutant libraries in vitro", pp. 63-68., 1988.
Landt, C., et al., Gene, vol. 96, "A general method for rapid site-directed mutagenesis using the polymerase chain reaction", pp. 125-128, 1990.
Kuipers, O. P., et al., Nucleic Acids Research, vol. 19, "Improved site-directed mutagenesis using PCR", p. 4556, 1991.
Zhou, Y., et al., Nucleic Acids Research, vol. 19, "Random mutagenesis of gene-sized DNA molecules by use of PCR with Taq DNA polymerase", p. 6052, 1991.
Zoller, M. J., Current Opinion in Biotechnology, vol. 2, "New molecular biology methods for protein engineering", pp. 526-531, 1991.
Caldwell, R. C., et al., PCR Methods and Applications, vol. 2, "Randomization of genes by PCR mutagenesis", pp. 28-33, 1992.
Sood, A. K., et al., 1981, Proceedings of the National Academy & Sciences, U.S.A., 78(1): 616-620.
Cullen, D., et al., 1987, Bio/Technology, 5: 369-376.
Matsuda, G., et al., 1981, FEBS Letters, 126(1): 111-113.
Karimi, S., and Ward, O. P., 1989, Journal of Industrial Microbiology, 4(3): 173-180.
Poutanen, K., 1988, Journal of Biotechnology, 7(4):271-282.
Kelly, M. A., et al., 19
Andreoli Peter Michael
Bakhuis Janna Gardina
Coutel Yves
De Graaff Leendert Hendrick
Der Veen Peter Van
Gist-brocades, N.V.
Moore W.
Wax Robert A.
LandOfFree
Cloning and expression of DNA molecules encoding arabinan-degrad does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Cloning and expression of DNA molecules encoding arabinan-degrad, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Cloning and expression of DNA molecules encoding arabinan-degrad will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1449798