EP0674717A1 - METHOD OF PRODUCING $g(a),$g(a)-TREHALOSE - Google Patents

METHOD OF PRODUCING $g(a),$g(a)-TREHALOSE

Info

Publication number
EP0674717A1
EP0674717A1 EP94926769A EP94926769A EP0674717A1 EP 0674717 A1 EP0674717 A1 EP 0674717A1 EP 94926769 A EP94926769 A EP 94926769A EP 94926769 A EP94926769 A EP 94926769A EP 0674717 A1 EP0674717 A1 EP 0674717A1
Authority
EP
European Patent Office
Prior art keywords
trehalose
membrane
ethanol
microfiltration
ultrafiltration
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP94926769A
Other languages
German (de)
English (en)
French (fr)
Inventor
Miroslav Bleha
Jaroslav Prchlik
Vladimir Janousek
Jan Schauer
Jiri Stuchlik
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
USTAV MAKROMOLEKULARNI CHEMIE AKADEMIE VED CESKE REPUBLIKY
VUFB AS
Original Assignee
USTAV MAKROMOLEKULARNI CHEMIE AKADEMIE VED CESKE REPUBLIKY
VUFB AS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by USTAV MAKROMOLEKULARNI CHEMIE AKADEMIE VED CESKE REPUBLIKY, VUFB AS filed Critical USTAV MAKROMOLEKULARNI CHEMIE AKADEMIE VED CESKE REPUBLIKY
Publication of EP0674717A1 publication Critical patent/EP0674717A1/en
Withdrawn legal-status Critical Current

Links

Classifications

    • 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
    • C12P19/12—Disaccharides

Definitions

  • the invention pertains to the method for producing disaccharide ⁇ , ⁇ -trehalose ( ⁇ -D-glucopyranosyl- ⁇ -D-glucopy- ranoside) , which is an important raw material or intermediate in food industry, pharmacy and biochemistry.
  • ⁇ , ⁇ -Trehalose may be obtained by isolation from a broad selection of natural substances, above all from Selaginella lepidophyla by origin from southwest of USA which is now grown in the countries of Middle East.
  • the most suitable source of this disaccharide is baker's yeast, above all for its good accessibility, low price and a relatively high content of a , ⁇ -trehalose.
  • the isolation of ⁇ , ⁇ -trehalose from baker's yeast was described for the first time already in 1936 (K. Myrback, B. Ortenblad: Biochem. Z . 288, 329 (1936)) and its modified form (Z.C. Stewart, N.K. icht yer, C.S. Hudson: J.Am.Cheia. Soc . 72, 2059 (1950)) is employed for the preparation of ⁇ , ⁇ -treh- alose up to the present time.
  • the object of the present invention is a method for producing crystalline ⁇ , ⁇ -trehalose by isolation from baker's yeast, which method consists in the extraction of baker's yeast with ethanol of concentration 70 to 96 volume %, vacuum concentration of the obtained extract to the content of ⁇ , ⁇ -trehalose 2 to 30 weight %, after removing the solid portion by filtration, and in the following membrane filtra ⁇ tion process consisting of ultrafiltration through an ultrafiltration membrane with the separation limit of molecular masses higher than 500 and, if desirable, of the preceding microfiltration through a microfiltration membrane with the mean pore diameter ranging from 0.05 to 5 ⁇ m, preferably from 0.05 to 0.15 ⁇ , where the obtained clear solution, after decolourizing with activated charcoal, is brought into contact with an ion exchanger in H + cycle and then with an ion exchanger in OH " cycle and the resulting demineralized solution is evaporated in vacuum to the concentration of 40 to 85 weight % and ⁇ , ⁇ -trehalose, crystallizing after
  • Another object of the method according to the inventa- tion consists in the fact that the continuous ultrafiltration is carried out through an ultrafiltration membrane with the separation limit of molecular masses higher than 500 and, if desirable, the preceding microfiltration is carried out through a microfiltration membrane with mean pore diameter ranging from 0.05 to 5 ⁇ m, preferably from 0.05 to 0.15 ⁇ m.
  • a styrene-divinylbenzene copolymer with strongly acid sulfo groups as a cation exchanger and a styrene- divinylbenzene copolymer with quaternary dimethy- lhydroxyethyl- ammonium exchanging groups as an anion exchanger.
  • the method according to the invention may be realized in such a way, that baker's yeast is extracted at ambient tempe ⁇ rature with 96% ethanol denatured with 2-propanol, the extracted yeast is filtered off and ethanol is removed from the filtrate by vacuum distillation.
  • the obtained aqueous or water- ethanolic solution with the concentration of ⁇ , ⁇ -treh ⁇ alose 10 to 40 weight % is subjected to ultrafiltration through an ultrafiltration membrane with the separation limit of molar masses higher than 500, or may be first subjected to microfiltration through a microfiltration membrane with the mean pore diameter ranging from 0.05 to 5 ⁇ m and then to ultrafiltration through an ultrafiltration membrane with the separation limit of molar masses higher than 500.
  • the clear aqueous solution after decolourizing with activated charcoal is deionized on the columns of cation exchanger and anion exchanger and concentrated in a vacuum evaporator to aqeous syrup with the concentration of ⁇ , ⁇ -trehalose 50 to 70 weight %.
  • the disaccharide crystallizes at normal or suppressed temperature.
  • Crystalline a, ⁇ -trehalose is isolated by filtration and dried at 40 to 60°C at reduced presure. If crystalline ⁇ , ⁇ -trehalose of purity higher than 99 weight % is produced, the crude substance has to be recrystaUized from ethanol after deionization realized in a similar way.
  • the method according to the invention renders ⁇ , ⁇ -tre ⁇ halose of high quality in a satisfactory yield, thus enabling its further direct use, e.g., in pharmacy or in food produc ⁇ tion.
  • Baker's yeast 1000 g is extracted under intense stirring at ambient temperature with 3000 ml 96 % ethanol denatured with 6 % 2-propanol for 45 min. The suspension is filtered and the filtration cake is washed with 250 ml ethanol and is further used for the isolation of ergosterol. The resulting filtrate is concentrated in a vacuum evaporator (20 mm Hg, bath temperature 60°C) to the volume of 400 ml and transferred into a filtration cell of capacity 450 ml provided with an efficient stirring of filtered liquid and microfiltration membrane Synpor" with the mean pore diameter 0.12 ⁇ m.
  • Overpressure 0.08 MPa is exerted above the filtered liquid and membrane by nitrogen introduced from a pressure cylinder and 380 ml liquid is filtered. This filtrate is subjected to ultrafiltration in the same cell provided with an ultrafiltratiom membrane from polysulfone having the separation limit of molecular masses 20,000. In this operati ⁇ on, overpressure above the filtered liquid and membrane is 0.25 MPa and 360 ml filtrate is obtained.
  • aqueous solution (360 ml) is decolourized by addition of 10 g activated charcoal, which is then filtered off, and the filtrate is deionized in a column of 300 ml Wofatit KPS in H + cycle and a column of 215 ml Wofatit SBK in OH " cycle.
  • the ion exchangers are washed with 1500 ml water and the combined solutions are evaporated in vacuum (20 mm Hg) to aqeous syrup containing 71 weight % ⁇ , -trehalose. After addition of 50 ml ethanol, crystallization proceeds at ambient temperature for 16 hours.
  • Crystalline ⁇ , ⁇ -trehalose is isolated by filtration of a fritted-glass filter G-2, the filtration cake is washed with 50 ml ethanol and dried in vacuum (50°C, 15 mm Hg) giving 31.5 g crystalline ⁇ , ⁇ -trehal ⁇ ose of purity 97.1 %. Another 2.7 g of the crystalline product is obtained by concentrating mother liquors and crystallization under above specified conditions.
  • Dried baker's yeast 250 g is mixed at laboratory temperature with 580 ml demineralized water to a paste which is then extracted with 2000 ml 96 % ethanol denatured with 7 % 2-propanol for 30 minutes. The suspension is filtered and the filtration cake is washed with 2150 ml of 70 % ethanol denatured with 2-propanol.
  • the filtrate obtained is concen ⁇ trated in a vacuum evaporator (20 mm Hg, temperature of water bath 60°C) to the volume of 400 ml, transferred into filtra- tion cell of capacity 450 ml with the continuous flow of filtered liquid over a membrane and with the ultrafiltration membrane from cellulose acetate having the separation limit of molecular masses 5000.

Landscapes

  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Microbiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Separation Using Semi-Permeable Membranes (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
EP94926769A 1993-09-28 1994-09-23 METHOD OF PRODUCING $g(a),$g(a)-TREHALOSE Withdrawn EP0674717A1 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
CZ932025A CZ281723B6 (cs) 1993-09-28 1993-09-28 Způsob izolace krystalické alfa, alfa-trehalosy
CZ202593 1993-09-28
PCT/CZ1994/000022 WO1995009243A1 (en) 1993-09-28 1994-09-23 METHOD OF PRODUCING α,α-TREHALOSE

Publications (1)

Publication Number Publication Date
EP0674717A1 true EP0674717A1 (en) 1995-10-04

Family

ID=5464225

Family Applications (1)

Application Number Title Priority Date Filing Date
EP94926769A Withdrawn EP0674717A1 (en) 1993-09-28 1994-09-23 METHOD OF PRODUCING $g(a),$g(a)-TREHALOSE

Country Status (6)

Country Link
EP (1) EP0674717A1 (cs)
JP (1) JPH08506248A (cs)
CA (1) CA2150256A1 (cs)
CZ (1) CZ281723B6 (cs)
SK (1) SK279610B6 (cs)
WO (1) WO1995009243A1 (cs)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1053670C (zh) * 1997-09-18 2000-06-21 大连理工大学 海藻糖的生产方法
WO2013042587A1 (ja) 2011-09-21 2013-03-28 株式会社林原 α，α－トレハロース二含水結晶含有粉末の製造方法
CN102504040A (zh) * 2011-11-16 2012-06-20 华南理工大学 一种从生产甾醇后的废弃酵母中提取海藻糖的方法
EP3744853A1 (en) * 2019-05-29 2020-12-02 Ohly GmbH Trehalose-rich yeast hydrolysate

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH04360692A (ja) * 1991-06-07 1992-12-14 Kanegafuchi Chem Ind Co Ltd トレハロースの製造方法
JPH0591890A (ja) * 1991-08-27 1993-04-16 Kanji Matsumoto トレハロースの製造方法
JPH05292986A (ja) * 1992-02-21 1993-11-09 Takeda Chem Ind Ltd トレハロースの製造法
JPH06145186A (ja) * 1992-11-05 1994-05-24 Nippon Shokuhin Kako Co Ltd α，α−トレハロースの製造法
BR9400368A (pt) * 1993-02-02 1994-08-23 Ajinomoto Kk Processo para isolamento e purificação de trehalose

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9509243A1 *

Also Published As

Publication number Publication date
JPH08506248A (ja) 1996-07-09
SK279610B6 (sk) 1999-01-11
CZ281723B6 (cs) 1996-12-11
CA2150256A1 (en) 1995-04-06
SK69795A3 (en) 1996-05-08
CZ202593A3 (en) 1996-10-16
WO1995009243A1 (en) 1995-04-06

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