Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical
Reexamination Certificate
1997-04-29
2001-06-05
Prats, Francisco (Department: 1651)
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound containing saccharide radical
C435S101000, C435S072000, C435S098000
Reexamination Certificate
active
06242224
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to a novel maltohexaose and maltoheptaose-forming amylase, and preparation and uses thereof, more particularly, to a maltohexaose and maltoheptaose-forming amylase which mainly forms maltohexaose and maltoheptaose from starch (designated as “AMYLASE” hereinafter) and preparation thereof, as well as microorganisms which produce such an amylase, saccharide compositions containing maltohexaose and/or maltoheptaose produced therewith or those containing maltohexaitol and/or maltoheptaitol obtainable by hydrogenating the maltohexaose and maltoheptaose, and compositions containing these saccharides.
2. Description of the Prior Art
Methods for specifically producing maltooligosaccharides such as maltose and maltotetraose by allowing specific amylases to act on starch have been employed on an industrial scale, and widely used in processes of compositions such as foods and pharmaceuticals. Demands for saccharides containing a relatively-large quantity of maltooligosaccharides such as maltohexaose and maltoheptaose have increased because, among maltooligosaccharides, those with a relatively-low molecular weight are considerably-low in sweetness, readily digestible and absorbable.
Enzymes or amylases, derived from microorganisms, which produce a relatively-large amount of maltohexaose when act on starch are known, but no amylase which forms maltooligosaccharides higher than maltoheptaose has been known.
Amylases which mainly produce maltohexaose from starch can be roughly classified into two groups based on their actions. The one is an exo-type maltohexaose-forming amylase which hydrolyzes amylaceous substances so as to remove successive maltohexaose units from the non-reducing chain ends, namely, maltohexaohydrolase (EC 3.2.1.98), and the other is an endo-type maltohexaose-forming amylase, namely, endo-type &agr;-amylase (EC 3.2.1.1) which acts on internal amylaceous saccharide chains to produce maltohexaose in a relatively-large quantity.
Kainuma et al. reported in
FEBS Letters
, Vol.26, pp.281-285 (1972) a microorganism of the species
Aerobacter aerogenes
which intracellularly forms an exo-type maltohexaose-forming amylase which is characteristic of its relatively-low optimum temperature and thermal stability, but it is insufficient in heat tolerance for an industrial use.
J. F. Kennedy et al. reported in
Starch
, Vol.31, pp.235-241 (1979) a microorganism of the species
Bacillus subtilis
forms an endo-type maltohexaose-forming amylase, and Takasaki reported in
Agricultural and Biological Chemistry
, Vol.46, pp.1539-1547 (1982) a microorganisms of
Bacillus circulans
G-6 forms such an amylase. Taniguchi reported in
Denpun Kagaku
(Starch Science), Vol.29, pp.107-116 (1982) a microorganism of the species
Bacillus circulans
F-2 forms such an amylase, and Hayashi et al. reported in
Agricultural and Biological Chemistry
, Vol.52, pp.443-448 (1988) a microorganism of the species
Bacillus
sp. H-167 forms such an amylase. It has been known that, among these amylases produced from such microorganisms, those from
Bacillus circulans
G-6
, Bacillus circulans
F-2 and
Bacillus
sp. H-167 form maltohexaose from starch in the maximum yield of about 25-30 w/w %, on a dry solid basis (d.s.b.), (the wording “w/w %” is designated as “%” hereinafter, if not specified otherwise), but they do not form maltoheptaose, and further they hydrolyze maltohexaose into maltose and maltotetraose as their enzymatic reactions proceed.
Now referring to amylases derived from microorganisms of the species
Bacillus subtilis
, they only form about 25% maltohexaose from starch and do not hydrolyze maltohexaose. Such amylases are not suitable for an industrial-scale production of high maltohexaose content saccharides because they do not form maltohexaose in a satisfactorily-high yield while forming a relatively-large quantity of lower molecular oligosaccharides under starch saccharification conditions, and because they could not be readily obtained in quantity from microorganisms.
In
Agricultural and Biological Chemistry
, Vol.42, pp.259-267 (1978) it was reported that a purified &agr;-amylase, prepared from malts, mainly forms maltohexaose and maltoheptaose during the early stage of the enzymatic reaction on starch. It is also reported that the formed maltohexaose and maltoheptaose are decomposed almost completely into lower molecular oligosaccharides such as maltose and maltotetraose as the enzymatic reaction proceeds, and, therefore, it scarcely produces saccharified products, mainly containing maltohexaose and maltoheptaose, from starch.
SUMMARY OF THE INVENTION
The present inventors pursued a novel amylase which mainly forms maltohexaose and maltoheptaose from starch and screened microorganisms which form such an amylase. As a result, they found that a novel microorganism of the genus Alcaligenes or Alcaligenes latus D2271, isolated from a soil nearby Komagatake in Yamanashi, Japan, forms a novel amylase which forms a relatively-large quantity of maltohexaose and maltoheptaose from starch (sometimes designated as “AMYLASE” in the specification), established the preparation of saccharide compositions containing maltohexaose and/or maltoheptaose by allowing the amylase to act on amylaceous substances, and also established compositions such as food products, cosmetics, pharmaceuticals and shaped products. Thus, they accomplished this invention.
REFERENCES:
patent: 5266467 (1993-11-01), Inglett
patent: 2001075 (1979-01-01), None
patent: 2038182 (1980-07-01), None
patent: 0210597 (1992-07-01), None
patent: 0038265 (1993-02-01), None
patent: 8910970 (1989-11-01), None
Lloyd et al, Chapter XXI ofStarch: Chemistry and Technology, Whistler et al, eds., 1984, pp. 617-623.*
Hayashi et al, “Production and Purification of New Maltohexaose-forming Amylases from Alkalophilic Bacillus sp. H-167”;Agricultural and Biological Chemistry, 52:443-448 (1988).
Kainuma et al, “Isolation and Action Pattern of Maltohexaose Producing Amylase from Aerobacter Aerogenes”;FEBS Letter, 26:281-285 (1972).
Kennedy et al, Action Pattern and Substrate Specificity of alpha-Amylase K, a Novel Aylase from a Strain of Bacillus Subtilis;Starch, 31:235-241 (1979).
Maeda et al, “Purification of R-enzyme from Malted Barley and Its Role in in vitro Digestion of Barley Starch Granules”;J. Jap. Soc. Starch Science, 26:117-127 (1979).
Maeda et al, “Digestion of Barley Starch Granules by the Combined Action of alpha- and beta-Amylases Purified fro mBarley and Barley Malt”;Agricultural and Biological Chemistry, 42:259-267 (1978).
Takasaki, Y., “Production of Maltohexaose by alpha-Amylase from Bacillus circulans G-6”,Agricultural and Biological Chemistry, 46:1539-1547 (1982).
Taniguchi et al, “Enzymic Digestion of Potato Starch Granules”;J. Jap. Soc. Starch Science, 29:107-116 (1982).
ATCCCatalogue of Bacteria&Bacteriophates, 17th Ed., pp. 13-14 (1989).
I. Maeda et al., “Digestion of Barley Starch Granules by the Combined Action of Alpha- and Beta-Amylases Purifird Barley and Barley Malt”, Agricultural Biological Chemical, vol. 42, pp. 259-267, 1978.
Y. Takasaki et al., “Production of Maltohexaose by Alpha-Amylase from Bacillus Circulans G-6”, Agricultural Biological Chemical, vol. 46, pp. 1539-1547, 1982.
R. -Q. He et al., “An asynchronous unfolding among molecular different regions of lobster D-glyceraldehyde-3-phosphate dehydrogenase and maltotetrsose- forming amylase from an Alcaligenes sp. during guanidine denaturation”, Biochimica et Biophysica Acta, vol. 1163, pp. 315-320, 1993.
CA 119: 27066p, A. Totsuka et al., “Maltooligosaccharide Compositions for Use as Sweetening Agents and Their Preparation from Starch with &agr;-Amylase and Debranching Enzymes”, Chemical Abstracts, vol. 119, No. 3, p. 77, Jul. 19, 1993.
CA 119: 48070a, A. Totsuka et al., “Reduced Starch Sugar Compositions Containing Maltohexitol and Meltoheptitol for use in Foods and Beverages”, Chemical Abstracts, vol. 119, No. 5, Aug. 2, 1993.
Chaen Hiroto
Miyake Toshio
Nakano Masayuki
Sugimoto Toshiyuki
Browdy & Neimark
Kabushiki Kaisha Mayashibara Seibutsu Kagaku Kenkyujo
Prats Francisco
LandOfFree
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