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US 5,858,748 AGranted
Gram-positive alkaliphilic microorganisms
AssigneeGenencor International, Inc.
InventorsBrian Edward Jones, William Duncan Grant, Nadine Claire Collins
Life of this patent
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Abstract
The present invention provides novel aerobic, Gram-positive alkaliphilic bacteria which have been isolated from in and around alkaline soda lakes. These alkaliphiles have been analyzed according to the principles of numerical taxonomy with respect to each other and also to a collection of known bacteria. In addition, these bacterial taxa are further circumscribed by an analysis of the lipid components which serve as chemotaxonomic markers. The alkaliphiles of the present invention produce alkalitolerant enzymes which are capable of performing their functions at high pH which makes them uniquely suited for applications requiring such extreme conditions.
Claims21
A method to prepare an alkali-tolerant enzyme wherein the method comprises
- separating bacteria which have been cultured in a suitable culture medium and recovering enzyme activity from said culture medium,
- wherein said bacteria consist of aerobic, Gram-positive, coccoid, obligate alkaliphilic bacteria having the following characteristics:
- a) forms orange-colored, circular colonies;
- b) grows optimally at about pH 10;
- c) gives a positive response to the following tests:
- 1) Gelatin hydrolysis
- 2) Penicillin G
- 3) Methicillin
- 4) Bacitracin;
- d) gives a negative response to the following tests:
- 1) N-acetylglucosamine
- 2) D-saccharose
- 3) Salicin
- 4) D-melibiose
- 5) Propionate
- 6) Glycogen
- 7) 3-hydroxybutyrate
- 8) Serine
- 9) Arginine
- 10) Methionine
- 11) Valine
- 12) Cellobiose.
19 more elements
A method to prepare an alkali-tolerant enzyme wherein the method comprises
- separating bacteria which have been cultured in a suitable culture medium and recovering enzyme activity from said culture medium,
- wherein said bacteria consist of aerobic, Gram-positive, short, irregular, rod-shaped, obligate alkaliphilic bacteria having the following characteristics:
- a) forms cream-yellow, small, circular colonies;
- b) grows optimally above pH 8;
- c) gives a positive response to the following tests:
- 1) Gelatin hydrolysis
- 2) Galactose
- 3) .beta.-glucosidase
- 4) Arginine
- 5) Bacitracin
- 6) N-acetyl-D-glucosamine
- 7) Cellobiose
- 8) Inosine
- 9) Uridine;
- d) gives a negative response to the following tests:
- 1) Maltose
- 2) Acetate
- 3) D-glucose
- 4) D-melibiose
- 5) Propionate
- 6) Valerate
- 7) Glycogen
- 8) Serine
- 9) Ampicillin
- 10) Penicillin G
- 11) Methicillin
- 12) Tetracyclin
- 13) Methyl pyruvate
- 14) Mono-methylsuccinate
- 15) .alpha.-ketobutyric acid.
27 more elements
A method to prepare an alkali-tolerant enzyme wherein the method comprises
- separating bacteria which have been cultured in a suitable culture medium and recovering enzyme activity from said culture medium,
- wherein said bacteria consist of aerobic, Gram-positive, coryneform, obligate alkaliphilic bacteria having the following characteristics:
- a) mature colonies are glistening bright orange/red;
- b) grows optimally between about pH 9 and 10;
- c) gives a positive response to the following tests:
- 1) Arginine
- 2) Methionine
- 3) Glycine
- 4) Valine
- 5) Methicillin
- 6) Tetracyclin
- 7) Bacitracin
- 8) .alpha.-cyclodextrin
- 9) Tween 40
- 10) Methyl pyruvate
- 11) Mono-methylsuccinate
- 12) .beta.-hydroxybutyric acid
- 13) Bromo-succinic acid;
- d) gives a negative response to the following tests:
- 1) Gelatin hydrolysis
- 2) Fumarate
- 3) Fructose
- 4) Galactose
- 5) D-saccharose
- 6) D-melibiose
- 7) Glycogen
- 8) L-serine
- 9) .beta.-glucosidase
- 10) N-acetyl-D-glucosamine
- 11) Cellobiose
- 12) D-sorbitol
- 13) D-trehalose
- 14) Turanose.
30 more elements
A method to prepare an alkali-tolerant enzyme wherein the method comprises
- separating bacteria which have been cultured in a suitable culture medium and recovering enzyme activity from said culture medium,
- wherein said bacteria consist of aerobic, Gram-positive, coccoid or short rod-shaped, obligate alkaliphilic bacteria consisting essentially of the following characteristics:
- a) cells are often found in pairs, exhibiting a characteristic snapping form of cell division so as to give a V-form arrangement;
- b) no growth below pH 8;
- c) on alkaline-agar, forms opaque, orange colored, punctiform or circular colonies;
- d) in alkaline-broth, growth (37.degree. C.) is flocculent, with the formation of a sediment and surface ring;
- e) grows optimally at above 30.degree. C., no growth above 40.degree. C.;
- f) grows at NaCl concentration of between 0-10%, no growth at a concentration of 12% NaCl;
- g) grows on yeast extract, peptones and carbohydrates;
- h) grows a positive response to the catalase reaction test;
- i) gives a negative response to the following tests:
- 1) KOH
- 2) Aminopeptidase
- 3) Oxidase
- 4) Gelatin hydrolysis
- 5) Starch hydrolysis.
13 more elements
A method to prepare an alkali-tolerant enzyme wherein the method comprises
- separating bacteria which have been cultured in a suitable culture medium and recovering enzyme activity from said culture medium,
- wherein said bacteria consist of aerobic, Gram-positive, straight or slightly curved rod-shaped obligate alkaliphilic bacteria consisting essentially of the following characteristics:
- a) cells are sometimes found in pairs, exhibiting a characteristic snapping for of cell division so as to give a V-form arrangement;
- b) no growth below pH 8;
- c) on alkaline-agar forms opaque yellow/ochre-colored circular colonies;
- d) grows between 10.degree. C. and 40.degree. C., no growth at 45.degree. C.;
- e) grows at NaCl concentration of 0-12%, no growth at NaCl concentration of 15%;
- f) grows on yeast extract;
- g) gives a positive response to the following tests:
- 1) Catalase reaction
- 2) Gelatin hydrolysis;
- h) gives a negative response to the following tests:
- 1) KOH
- 2) Aminopeptidase
- 3) Oxidase
- 4) Starch hydrolysis.
13 more elements
A method to prepare an alkali-tolerant enzyme wherein the method comprises
- separating bacteria which have been cultured in a suitable culture medium and recovering enzyme activity from said culture medium,
- wherein said bacteria consist of aerobic, Gram-positive, straight or slightly curved rod-shaped, obligate alkaliphilic bacteria consisting essentially of the following characteristics:
- a) cells are often found in pairs, sometimes in short chains of 2 to 4 cells;
- b) no growth below pH 7.5;
- c) on alkaline-agar, forms circular, cream-colored colonies;
- d) grows between 10.degree. C. and 45.degree. C., no growth at 50.degree. C.;
- e) grows at NaCl concentration of 0-15%;
- f) gives a positive response to the following tests:
- 1) Catalase reaction
- 2) Oxidase reaction
- 3) Gelatin hydrolysis
- 4) Starch hydrolysis;
- d) gives a negative response to the following tests:
- 1) KOH
- 2) Aminopeptidase.
12 more elements
A method to prepare an alkali-tolerant enzyme wherein the method comprises
- separating bacteria which have been cultured in a suitable culture medium and recovering enzyme activity from said culture medium,
- wherein said bacteria consist of aerobic, Gram-positive, obligate alkaliphilic bacterium consisting essentially of the following characteristics:
- a) cells are initially spherical, developing into short rods, exhibiting a characteristic snapping form of cell division so as to give a V-form arrangement;
- b) no growth below pH 8;
- c) on alkaline-agar, forms circular, convex, entire, opaque, orange to deep salmon pink colonies;
- d) grows at 10.degree. C. to 37.degree. C., no growth at 40.degree. C.;
- e) grows on yeast extract;
- f) grows at NaCl concentration of 0-12%;
- g) gives a positive response to the following tests:
- 1) Catalase reaction
- 2) Aminopeptidase
- 3) Gelatin hydrolysis;
- h) Gives a negative response to the following tests:
- 1) KOH
- 2) Oxidase reaction
- 3) Starch hydrolysis.
13 more elements
Dependencies are inferred from the claim text.