Enzymes with aminopeptidase activity

Details Enzymes with aminopeptidase activity Enzymes with aminopeptidase activity

2000-11-22

2002-07-02

Nashed, Nashaat T. (Department: 1652)

Food or edible material: processes, compositions, and products

Fermentation processes

Of farinaceous cereal or cereal material

C426S549000, C435S212000, C435S252330, C536S023200

Reexamination Certificate

active

06413559

ABSTRACT:

FIELD OF THE INVENTION
The present invention relates to an enzyme exhibiting aminopeptidase activity, a method for producing said enzyme, an enzyme preparation containing said enzyme exhibiting aminopeptidase activity, and use of said enzyme for various industrial purposes.
BACKGROUND OF THE INVENTION
Protein hydrolysates are being used in numerous food products. Traditionally protein hydrolysates were produced by acid hydrolysis, but today enzymatic hydrolysis is regarded as an attractive alternative.
One of the main problems of protein hydrolysates is that they often taste bitter. When using e.g. soy protein or casein, which are rich in hydrophobic L-amino acids, as the protein source, the protein hydrolysate tends to have bitter taste. In general it is believed that whether the taste of proteins is bitter or not depends on the average hydrophobicity of the L-amino acid residues, such as valine, leucine, isoleucine, phenylalanine, tyrosine and tryptophan.
A vast number of enzymes exhibiting peptidase activity are capable of performing enzymatic hydrolysis on vegetable, yeast and/or animal proteins, resulting in highly nutritious protein hydrolysates useful as food additives in products such as soups, sauces, gravies, paste, tofu, bouillon, seasonings, baby formulas, snacks, ready-to-eat meals etc.
Peptidases
All peptidases or proteases are hydrolases which act on proteins or its partial hydrolysate to decompose the peptide bond.
EP 427,385 (The Japanese R & D Association) discloses a genomic gene encoding an alkaline protease derived from yellow molds such as
Aspergillus oryzae.
JP-0-2002374 and JP-0-2002375 (Shokuhin), describes an alkaline protease derived from
Aspergillus oryzae
for use in medicine, food, and detergents.
SU-891777 (Khark) concerns a microbial protease from
Aspergillus oryzae,
which can be used in food, medicine etc.
JP-5-4035283 (Ajinomoto K K) discloses preparation of enzymes from e.g.
Aspergillus oryzae
exhibiting endopeptidase activity, which can hydrolyse proteins almost completely.
WO 94/25580 (Novo Nordisk A/S) describes a method for hydrolysing vegetable or animal protein by incubating with a proteolytic enzyme preparation derived from a strain of
Aspergillus oryzae.
Aminopeptidases
A subgroup of peptidases (proteases) are called aminopeptidases and are classified under the Enzyme Classification number E.C. 3.4.11 (aminopeptidases) in accordance with the Recommendations (1992) of the International Union of Biochemistry and Molecular Biology (IUBMB)).
Aminopeptidases are capable of removing one or more amino terminal residues from polypeptides.
JP-7-5034631 (Noda) discloses a leucine aminopeptidase derived from yellow koji mold, which includes
Aspergillus oryzae.
JP-7-4021798 (Zaidan Hojin Noda Sangyo) describes the production of miso by adding of a leucine aminopeptidase II prepared by cultivating a number of molds, including
Aspergillus oryzae
strain 460 and strain IAM 2616.
Van Heeke et al., Bioch. Biophys. Acta, (1992), 1131, 337-340, have disclosed the cloning of a 30 kDa aminopeptidase from the bacteria
Vibrio proteolyticus
deposited at the American Type Culture Collection under the ATCC No. 15338.
Aspergillus oryzae
460 is known to produce a number of leucine aminopeptidases. The molecular weight of three of these was calculated to 26,500, 56,000 and 61,000, respectively determined by gel filtration (Nakada et al., Agr. Biol. Chem, (1972), 37(4), 757-765; Nakada et al., Agr. Biol. Chem, (1972), 37(4), 767-774; Nakada et al., Agr. Biol. Chem, (1972), 37(4), 775-782). The
Aspergillus oryzae
460 strain is deposited at the American Type Culture Collection as
A. oryzae
(ATTC no. 20386).
Reduction of Bitter Taste of Protein Hydrolysates
EP 65,663 and EP 325,986 (Miles Inc.) concerns enzymatic hydrolysis of proteins using a mixture of enzymes containing
Aspergillis oryzae
derived proteases. The obtained protein hydrolysate has a bland, non-bitter taste.
JP-4-7029577 (Asahi Electro-chemical Co.) concerns a protease, derived from
Aspergillus oryzae,
which does not produce any bitter component when decomposing protein.
Prior art discloses a plethora of enzymes exhibiting peptidase, aminopeptidase and other enzyme activities. Said enzymes may be derived from a number of microorganisms, including the fungus species
Aspergillus oryzae.
In general products, useful for producing protein hydrolysates without a bitter taste, comprise a mixture of peptidase and aminopeptidase activities.
It would therefore be desirable to be able to provide a single-component enzyme (i.e. substantially without any side activity) exhibiting only an activity useful for reducing the bitterness of protein hydrolysates used in food products.


REFERENCES:
patent: 5112812 (1992-05-01), Samuelsson et al.
patent: 0 065 663 (1982-12-01), None
patent: 0 325 986 (1988-01-01), None
patent: 0 257 821 (1988-03-01), None
patent: WAO 94/26882 (1994-11-01), None
Matsuoka, H. et al., J. Agric. Food Chem., vol. 39, pp. 1392-1395 (1991).

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