WO2001068809A1 - Bacillus circulans b-65, cyclodextrine glucanotransferase obtenue a partir de bacillus circulans b-65 et utilisation pour la production de cyclodextrine - Google Patents

Bacillus circulans b-65, cyclodextrine glucanotransferase obtenue a partir de bacillus circulans b-65 et utilisation pour la production de cyclodextrine Download PDF

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Publication number
WO2001068809A1
WO2001068809A1 PCT/YU2000/000010 YU0000010W WO0168809A1 WO 2001068809 A1 WO2001068809 A1 WO 2001068809A1 YU 0000010 W YU0000010 W YU 0000010W WO 0168809 A1 WO0168809 A1 WO 0168809A1
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Prior art keywords
cyclodextrin
cgt
ase
marked
starch
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PCT/YU2000/000010
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English (en)
Inventor
Slobodan Stankovic
Dragan Pesic
Draga Simic
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Ad 'zdravlje'
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Priority to AU36334/00A priority Critical patent/AU3633400A/en
Priority to UA2002108074A priority patent/UA75352C2/uk
Publication of WO2001068809A1 publication Critical patent/WO2001068809A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1048—Glycosyltransferases (2.4)
    • C12N9/1051—Hexosyltransferases (2.4.1)
    • C12N9/1074—Cyclomaltodextrin glucanotransferase (2.4.1.19)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/18—Preparation of compounds containing saccharide radicals produced by the action of a glycosyl transferase, e.g. alpha-, beta- or gamma-cyclodextrins

Definitions

  • This invention belongs to the field of biotechnology and relates to a process for producing cyclodextrin using effect of cyclodextrin glucanotransferase (to be referred to as »CGT-ase « hereinafter) on starch and production of CGT-ase by bacterium Bacillus circulans B-65.
  • CGT-ase cyclodextrin glucanotransferase
  • the invention is classified into following classes: C08B 37/16; C12P 19/18; C12N 1/20; C12N 9/10.
  • This invention solves the problem of biosynthesis of CGT-ase which is conviniant for production of cyclodextrin, as well as the problem of production of cyclodextrin using effect of CGT-ase on starch.
  • Biosynthesis of CGT-ase is most often performed by bacteria belonging to the genus Bacillus which mostly produce CGT-ase which acting on starch forms the mixture of ⁇ -, ⁇ - and ⁇ -cyclodextrins in different ratio, while the yield of a single cyclodextrin is not high, and their separation raises the cost of production.
  • Biosynthesis of CGT-ase is, according to the invention, performed by novel alkalophilic bacterium Bacillus circulans B-65 characterised by production of a large quantity of CGT-ase which is conviniant for production of cyclodextrin. Cyclodextrins are produced using CGT-ase of B. circulans bacterium which is characterised by high degree of conversion of starch to cyclodextrins and is specific for ⁇ -cyclodextrin formation.
  • Cyclodextrins are produced by effect of CTG-ase on starch. In the process of cyclodextrins production the most important phase is production of CTG-ase.
  • CTG-asa is produced by some species of bacteria, among which the most frequent are the bacteria belonging to genus Bacillus: B. macerans, B. circulans, B. megaterium, B. coagulans, B. lentus, B. ohbensis, B. ⁇ rmus, B. amyloliquefaciens, B. stearothermophilus, B. subtilis, B. cerus.
  • CTG-ase is produced by some other bacteria, apart from genus Bacillus, for example: Klebsiella pneumoniae, Micrococcus luteus, Micrococcus varians.
  • the most often producers of CGT-ase are Bacillus macerans and alkalophilic bacteria belonging to genus Bacillus.
  • CGT-ase is produced by growing of bacteria in liquid medium under the aerobic conditions in fermentor. This process is significantly expensive, and bacteria produce certain quantity of CGT-ase. Expensive process for production of CGT-ase significantly influences the high cost of the final product ie. cyclodextrin, which restricts its broad usage in a number of industrial fields.
  • CGT-ase In the process of cyclodextrin production, the characteristics of CGT-ase, produced by bacteria, are also very important. The characteristics of CGT-ases, produced by different species of bacteria belonging to genus Bacillus are shown in the table 1. Table I. Characteristics of CGT-ases of different microorganisms
  • CGT-ases from different microorganisms, show similarities in characteristics, but, there are certain differences. CGT-ases show the greatest differences in specific qualities for ⁇ -, ⁇ - or ⁇ -cyclodextrin forming. CGT-ases mostly form the mixture of -, ⁇ - and ⁇ -cyclodextrins in different ratio. However, there are CGT-ases which do not form ⁇ - or ⁇ -cyclodextrins. Majority of CGT- ases form the mixture of cyclodextrins, where ⁇ -cyclodextrin prevails, while ⁇ -cyclodextrin is in the lowest amount.
  • Alkalophilic microorganisms produce CGT-ases which give the highest content of ⁇ -cyclodextrin, compared to ⁇ - and ⁇ -cyclodextrins.
  • CGT-ases of alkalophilic mocroorganisms form ⁇ -cyclodextrin which represents more than 70% of total cyclodextrins. From data given in table 1 we can conclude that CGT-ases, produced by strains belonging to species Bacillus macerans, give the highest yield of ⁇ -cyclodextrin compared to other cyclodextrins. Besides B. macerans, CGT-ase produced by B.
  • amyloliquefaciens AL 35 also forms ⁇ - cyclodextrin in yield up to 95%.
  • CGT-ase produced by Bacillus subtilis No. 313 forms ⁇ -cyclodextrin, but it does not form nither ⁇ - nor ⁇ -cyclodextrin.
  • CGT-ase of Bacillus sp. AL-6 gives the highest yield of ⁇ -cyclodextrin and ⁇ -cyclodextrin, but it does not form ⁇ -cyclodextrin.
  • Biosynthesis of CGT-ase which is later used for cyclodextrin production, is performed by alkalophilic bacterium Bacillus circulans B-65. Biosynhesis is performed by growing the bacterium in alkaline (pH 10) liquid medium, under aerobic conditions in fermentor. After finished biosynthesis bacterial cells are removed by centrifuging or filtering, and then, extracellularly produced CGT-ase is concetrated using ultrafiltration. By the effect of CGT-ase, produced such way, on starch, in concentration 5% to
  • Novelity of this invention is that biosynthesis of CGT-ase, which is used for production of cyc ⁇ odextrine, is performed using alkalophilic bacterium B. circulans B-65.
  • the bacterium was isolated from the soil taken from the area of Leskovac.
  • B. circulans was selected from around 2000 colonies of bacteria which were characterized by production of CGT-ase.
  • the isolate, which was later identified and marked as B. circulans B-65 showed considerably higher activity of production of CGT-ase, and that characteristic clearly distinguished it from other isolates. Consequently, alkalophilic B. circulans B-65 is a novel strain characterized by the fact that it produces great quantity of CGT-ase which can be used for production of cyclodextrins.
  • Bacterium Bacillus circulans B-65 is deposited with National Collection of Agricultural and Industrial Microorganisms (NCAIM) - Budapest, under following number NCAIM (P) 001277.
  • NCAIM National
  • CGT-ase which is produced by our alkalophilic strain B. circulans B-65 is applied for production of ⁇ -cyclodextrin.
  • CGT-ase of our strain is specific for ⁇ -cyclodextrin formation and is characterized by high degree of conversion of starch to ⁇ -cyclidextrin. Under described conditions of reaction, with concentration of starch of 5%, CGT-ase performs conversion of starch to cyclodextrins up to 45%. From total amount of produced cyclodextrins, the amount of ⁇ -cyclodextrin is nearly 95%, which represents higher percentage compared to data from available literature.
  • CGT-ase producing bacteria were isolated from 54 different samples of soil, which were taken from the area of Leskovac.
  • Soil suspension (0,5 to 1,0 g) in 10 ml of sterile, distillated water, was heated for 10 minutes at 80°C. After heating, 0,1 ml of soil suspension was spread on agar plates with Park medium containing: 1% soluble starch,
  • CGT-ase for further selection of isolates, was performed by growing of 45 selected alkalophilic isolates in liquid medium in Erlenmeyer-flasks with stirring. Incubation was performed at 37°C for 48 hours with continuous stirring 200 rpm. On the third and fourth day, samples were taken, and after removal of bacterial cells by centrifuging (10000 x g, 10 min, +4°C) amylolytic and cyclodextrinogenic activities of supernatant were determined.
  • Amylolytic activity of isolate was within the range from 0,4 U/ml to 12,17 U/ml.
  • the greatest number of isolates showed amylolytic activity from 1 U/ml to 2 U/ml.
  • the number of isolates which showed higher amylolytic activity was gradually reduced, so, amylolytic activity higher than 5 U/ml was shown only by 3 isolates.
  • Cyclodextrinogenic activity was within the range from 0,019 U/ml to 0,530 U/ml. Cyclodextrinogenic activity of the greatest number of isolates was up to 0,100 U/ml. Cyclodextrinogenic activity higher than 0,200 U/ml was showen only by one isolate.
  • Isolate B-65 showed amylolytic activity of 12,17 U/ml and cyclodextrinogenic activity of 0,530 U/ml, which were considerably higher compared to other isolates.
  • Morphological and taxonomic characteristics of bacterium For purpose of identification, morphological and physiological-biochemical characteristics of bacterium were examined. The results of examination were analysed using »Bergeys Manual of Systematic Bacteriology « (Sneath, 1968). Bacillus circulans (ATCC 4513) was used as a referent strain.
  • bacterium is: rod shaped, a size of 0,6-0,7 x 2 - 4 ⁇ m, motile, Gram positive, it forms ellipse shaped endospores, it grows under aerobic and anaerobic conditions and it produces catalase. These characteristics show that the bacterium belongs to the genus Bacillus.
  • the bacterium shows negative Voges-Proskauer reaction; it produces acids from glucose, arabinose, xylose, and mannite; it does not produce gas from glucose; it hydrolyses gelatin and starch; it does not use citrates; it does not produce indole; it does not grow in media with pH 6,8 and 5,7 and shows good growth in media with 2, 5, 7 and 10% NaCl.
  • Morphological and physiological-chemical characteristics of isolate B-65 are mostly identical with the ones of the species Bacillus circulans.
  • isolate B-65 shows good growth in alkaline medium (pH 10), and it does not grow in neutral medium, such as nutrient broth (pH 6,8), which differs it from described members of the species Bacillus circulans.
  • the isolate B-65 also differs from the members of the species Bacillus circulans by characteristic to grow in the medium with 10% NaCl.
  • isolate B-65 was identified as a member of the species Bacillus circulans. Because of its charasteristic to show good growth in alkaline medium (pH 10), and no growth in neutral medium, the isolate B-65 was marked as - alkalophilic Bacillus circulans B-65. The characteristics of isolate B-65 are shown in the table 2.
  • Biosynthesis of CGT-ase by B. circulans B-65 Biosynthesis of CGT-ase by B. circulans B-65 is carried out by growing in alkaline (pH 10) liquid medium.
  • Nutrient broth consists of: 1% soluble starch, 0,5% peptone- 1, 0,5% yeast extract, 0,1% K 2 HPO 4 , 0,02% MgSO 4 x 7H 2 0, 1% Na 2 CO 3 .
  • Inoculum for the process of CGT-ase production was prepared by inoculation of culture taken from agar slant into liquid medium. Incubation was carried out at 37°C with continuous stirring 200 rpm for 48 hours. Fermentor with fresh medium was inoculated with 5% inoculi.
  • CGT-ase In the course of cellular growth CGT-ase is produced. Until the phase of growth is over, amount of cyclodextrinogenic activity gets at 0,20 U/ml, which represents a bit more than 30%, compared to the value of activity obtained after 96 hours of growing. After finishing of the phase of growth, production of CGT-ase continues, so that after 96 hours of growing, cyclodextrinogenic activity gets up the value of 0,60 U/ml, and amylolytic activity 23,37 U/ml. Increase of cyclodextrinogenic and amylolytic activities after finishing the phase of growth, amounts to nearly 70% of total value. So, CGT-ase is more reachly produced by the isolate B-65, after the phase of cellular growth is over.
  • B. circulans B-65 amylolytic and cyclodextrinogenic activities increase rateably, which leads to the conclusion that B. circulans B-65 produces CGT-ase, and it does not produce any other amylolytic enzymes.
  • This conclusion is confirmed by determination of the contents of cyclodextrins and reducing sugars during incubation of CGT-ase of B. circulans B-65 with starch. The results showed that cyclodextrin formation occurred during reaction, while the amount of reducing sugars did not increase, which would have happened in the presence of other amylolytic enzymes.
  • CGT-ase biosynthesis by B. circulans B-65 compared to some other producers of this enzyme, is that the biosynthesis of CGT-ase by B. circulans B-65 is carried out in very alkaline medium (pH 10), while the risk of infection with some other microorganisms is reduced, which is very important for industrial conditions of production.
  • CGT-ase from B. circulans B-65 Characteristics of CGT-ase from B. circulans B-65
  • B. circulans B-65 produces CGT-ase in a great quantity
  • this CGT-ase is characterized by good properties concerning thermostability and optimum temperature for activity, and also concerning pH stability and optimum pH.
  • CGT-ase from B. circulans B-65 is characterized by the fact that it is stable at temperatures up to 55°C, and in the presence of Ca 2+ ion its stability increases up to 60°C. Also, the optimum temperature for its effect is 60°C. Thanks to the high optimum temperature of effect and high thermostability of CGT-ase from B. circulans B-65, process for production of cyclodextrin can be carried out at temperature of 60°C. The advantage of production of cyclodextrins at high temperature is that under these conditions of reaction, the risk of infection by microorganisms is reduced. Also, at higher temperatures the speed of enzyme reaction is higher which reduces duration of the process of cyclodextrin production.
  • CGT-ase from B. circulans B-65 is characterized by the fact that the optimum pH for its activity is in range from pH 5,0 to 6,0 - however CGT-ase retains its activity in quite wide range of pH in alkaline medium.
  • CGT-ase from B. circulans B-65 is stable in the range of pH 6,5 to 10,0. Thanks to theese properties of CGT-ase the production of cyclodextrin can be carried out at wide range of pH.
  • Table 4 Cyclodextrinogenic activity of CGT-ase at different pH values.
  • Relative activity is expressed as a percentage in relation to maximal activity obtained at pH 5,0
  • Obtaining of cyclodextrin was carried out by effect of CGT-ase from B. circulans B-65 on starch, with concentration in range from 5% to 15%. Thanks to high optimum temperature of effect and high thermostability of CGT-ase from B. circulans B-65, process for obtaining of cyclodextrin can be carried out at temperature up to 60°C. Concerning pH values, obtaining of cyclodextrin can be carried out in wide range from pH 6,5 to 10,0.
  • cyclodextrins After enzyme reaction for obtaining of cyclodextrin is over, they can be removed from reaction mixture by precipitation, using trichloroethylene or toluene, for example. From precipitated cyclodextrins, ⁇ -cyclodextrin can be removed by crystallization thanks to the fact that it is significantly less soluble in comparison with other cyclodextrins. Cyclodextrins obtained by precipitation are resolved in distillated water with heating at 100°C, and then crystallization of ⁇ -cyclodextrin by gradual reduction of temperature to 20 - 25°C with gentle stirring. The crystals of ⁇ -cyclodextrin are subsequently removed by filtration, washed by small quantity of cold water and dried at 70°C.
  • circulans B-65 is characterized by the fact that it is specific for formation of ⁇ -cyclodextrin and it converts high percentage of starch to ⁇ -cyclodextrin. Consequently, using CGT-ase from our strain has significant advantages for obtaining of ⁇ - cyclodextrin, in comparison with CGT-ases from other known bacteria.
  • Producing of CGT-ase was carried out by growing B. circulans B-65 in liquid medium containing: 1% soluble starch, 0,5% peptone, 0,5% yeast extract, 0,1% K 2 HPO 4 , 0,02% MgSO 4 x 7H 2 O, 1% Na 2 CO 3 .
  • Inoculum for the process of production of CGT-ase was prepared by inoculation of 450 ml of liquid Horikoshi II medium with the culture from agar slant. Incubation was being carried out for 48 hours at 37°C with continuous shaking 200 rpm. Fermentor containing 9 1 of Horikoshi II medium was inoculated with 450 ml of inoculum.
  • B. circulans B-65, grown such way, after 96 hours, extracellularly produced CGT- ase with amylolytic activity of 23,4 U/ml and cyclodextrinogenic activity of 0,6 U/ml. After concentration of enzyme by ultrafiltration, amylolytic activity of 450 U/ml and cyclodextrinogenic activity of 12 U/ml were obtained.
  • Soluble starch (300 g) was diluted in 6 1 of water with heating. Temperature was adjusted to 40°C and pH to 6,5. CGT-ase in concentration of 6U (cyclodextrinogenic activity )/g of starch, was added to starch solution. Incubation was being carried out for 30 hours at 40°C.
  • reaction was stopped by heating of reaction mixture for 10 minutes at 100°C. Reaction mixture was subsequently decolourized by activated charcoal. After removing of activated charcoal by filtration, cyclodextrins were precipitated from the reaction mixture (6 1) by trichloroethylene (1,2 1). Precipitated cyclodextrins were subsequently removed by filtration, and then dried at 70°C. Final yield was 107 g of cyclodextrins from 300 g of starch.

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Abstract

L'invention se rapporte à un procédé de production de cyclodextrine par une cyclodextrine glucanotransférase (EC 2.4.1.19.), produite de manière extracellulaire par une nouvelle bactérie alcalophile, Bacillus circulans B-65. La cyclodextrine glucanotransférase issue de Bacillus circulans B-65 se caractérise par un degré élevé de conversion de l'amidon en cyclodextrines et elle est spécifique de la formation de béta-cyclodextrines.
PCT/YU2000/000010 2000-03-14 2000-04-06 Bacillus circulans b-65, cyclodextrine glucanotransferase obtenue a partir de bacillus circulans b-65 et utilisation pour la production de cyclodextrine WO2001068809A1 (fr)

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AU36334/00A AU3633400A (en) 2000-03-14 2000-04-06 Bacillus circulans b-65, cyclodextrin glucanotransferase obtained therefrom and use to produce cyclodextrin
UA2002108074A UA75352C2 (en) 2000-03-14 2000-06-04 STRAIN OF BACTERIUM BACILLUS CIRCULANS B-65 û PRODUCER OF CYCLOMALTODEXTRINGLUKANOTRANSFERASE, METHOD FOR ISOLATION THEREOF AND USE FOR THE PREPARATION OF ?-CYCLODEXTRIN

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YUP015300 2000-03-14
YUP-153/00 2000-03-14

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100374555C (zh) * 2006-01-19 2008-03-12 山东大学 一种利用酵母生产β-环糊精的方法

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2556117C2 (ru) * 2013-12-05 2015-07-10 Федеральное государственное бюджетное научное учреждение "Всероссийский научно-исследовательский институт крахмалопродуктов" (ФГБНУ ВНИИК) СПОСОБ ПОЛУЧЕНИЯ β-ЦИКЛОДЕКСТРИНА

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4477568A (en) * 1981-09-24 1984-10-16 Proefstation voor Aardappelverweking-TNO en Cooperatieve Verkoop en Produktievereniging van Aardappelmeel en Derivaten AVERE B.A. Process for the manufacture of cyclodextrin
WO1989001044A1 (fr) * 1987-08-03 1989-02-09 Genetics Institute, Inc. Procede de preparation de cyclodextrines
WO1989003421A1 (fr) * 1987-10-15 1989-04-20 Novo Industri A/S Transferase glycosyliques de cyclodextrine thermostables, sa production et son utilisation
US5658390A (en) * 1994-06-29 1997-08-19 American Maize-Products Company Purification of beta cyclodextrin

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4477568A (en) * 1981-09-24 1984-10-16 Proefstation voor Aardappelverweking-TNO en Cooperatieve Verkoop en Produktievereniging van Aardappelmeel en Derivaten AVERE B.A. Process for the manufacture of cyclodextrin
WO1989001044A1 (fr) * 1987-08-03 1989-02-09 Genetics Institute, Inc. Procede de preparation de cyclodextrines
WO1989003421A1 (fr) * 1987-10-15 1989-04-20 Novo Industri A/S Transferase glycosyliques de cyclodextrine thermostables, sa production et son utilisation
US5658390A (en) * 1994-06-29 1997-08-19 American Maize-Products Company Purification of beta cyclodextrin

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
NITSCHKE L ET AL: "MOLECULAR CLONING, NUCLEOTIDE SEQUENCE AND EXPRESSION IN ESCHERICHIA COLI OF THE BETA-CYCLODEXTRIN GLYCOSYLTRANSFERASE GENE FROM BACILLUS CIRCULANS STRAIN NO. 8", APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,DE,SPRINGER VERLAG, BERLIN, vol. 33, no. 5, 1 August 1990 (1990-08-01), pages 542 - 546, XP000676776, ISSN: 0175-7598 *
RENDLEMAN JACOB A JR: "Enhancement of cyclodextrin production through use of debranching enzymes.", BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY, vol. 26, no. 1, 1997, pages 51 - 61, XP000990332, ISSN: 0885-4513 *
TOMITA KENJI ET AL: "Purification and properties of a cyclodextrin glucanotransferase from Bacillus autolyticus 11149 and selective formation of beta-cyclodextrin.", JOURNAL OF FERMENTATION AND BIOENGINEERING, vol. 75, no. 2, 1993, pages 89 - 92, XP000990285, ISSN: 0922-338X *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100374555C (zh) * 2006-01-19 2008-03-12 山东大学 一种利用酵母生产β-环糊精的方法

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