CN102504040A - Method for extracting trehalose from waste yeast after sterol production - Google Patents
Method for extracting trehalose from waste yeast after sterol production Download PDFInfo
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- CN102504040A CN102504040A CN201110362849XA CN201110362849A CN102504040A CN 102504040 A CN102504040 A CN 102504040A CN 201110362849X A CN201110362849X A CN 201110362849XA CN 201110362849 A CN201110362849 A CN 201110362849A CN 102504040 A CN102504040 A CN 102504040A
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Abstract
The invention discloses a method for extracting trehalose from waste yeast after sterol production. The method comprises the following steps: (1), waste yeast after sterol production is heated so as to remove remanent solvent and then dried into powder; (2), the yeast and alcohol are mixed and heated to 60 to 90 DEG C; the yeast is removed through centrifugation of extracting solution; (3), active carbon is added into the extracting solution after centrifugation in step (2), and under the condition that the PH value is 3.5 to 4.5 and the temperature is 25 to 35 DEG C, stirring is performed for decolourization; (5), the extracting solution processed in the step (3) flows through negative-positive or positive-negative, or positive-negative-positive ion resin tandem type exchange column so as to remove pigment and salt; (5), the extracting solution processed in the step (4) is condensed through decompression, the sugar concentration in the concentrated solution is controlled, then ethanol is added, and oscillation for shaking up is performed; and stewing after taking out is performed, and white crystal of trehalose is obtained. The method ensures that the waste yeast after sterol production can be used, and the economic value of the yeast is improved greatly.
Description
Technical field
The present invention relates to the microbial fermentation field, be specifically related to the metabolism of yeasts product and extract.
Background technology
Trehalose is a kind of irreducibility disaccharide, and its unique defencive function can make biomacromolecule (like protein, nucleic acid etc.) under severe environment such as external high temperature, dehydration, escape injury.The character of trehalose and performance make it have a wide range of applications at aspects such as food, healthcare products, medicine, makeup, agriculturals.Aspect food, trehalose can be used for preventing also can be used as the quality improver and the seasonings of some condiment, food because of dry or the freezing sex change that causes.Aspect healthcare products, trehalose is cooked the freeze-drying survival rate that lyophilized vaccine can improve bacterium greatly, and freeze-dried products is preserved at normal temperatures for a long time.Aspect medical, trehalose mainly is as the stablizer of reagent with the diagnosis medicine; Also can be used for toothpaste, medicine for oral administration, coated tablet etc. as sweeting agent, quality improver and stablizer, it have hypoglycemic, effect such as protect the liver, also can be made into verivate and be used for carcinostatic agent, antineoplastic agent.Aspect makeup, the moisture retention trehalose that has has been confirmed to be new cosmetic material, is used for the drying that skin cosmetics can suppress skin, also can be used for oral cavity freshener, oral cavity perfume compound etc.
The content of trehalose in yeast can reach 20%, and therefore traditional trehalose working method mainly is from yeast, to extract.Yeast generally as offal treatment, is easy to contaminate environment after with the non-polar solvent extract sterol, but this moment yeast itself cell walls pulverized, intravital various enzymes are inactivation also, the most of composition except that sterol also is retained in the yeast body.The present invention utilizes the discarded yeast behind the production sterol to be raw material, therefrom extracts trehalose, has both practiced thrift expense of raw materials, turns waste into wealth, and can reduce pollution again, the protection environment.
Summary of the invention
The objective of the invention is in order to develop the method for extraction trehalose a kind of discarded yeast after producing sterol.
The present invention realizes through following technical scheme:
Extract the method for trehalose a kind of discarded yeast after producing sterol, comprise the steps:
(1) the discarded yeast heating behind the production sterol is dried to powder again to remove residual solvent;
(2) be that 20～60% alcohol mixes with yeast and weight percent concentration, be heated to 60～90 ℃, lixiviate 30min～120min; The centrifugal yeast of removing of extracting solution, described yeast and alcohol weight ratio are 1: (10～30);
(3) adding activated carbon in the extracting solution after step (2) is centrifugal, is 3.5～4.5 at pH, under the condition that temperature is 25～35 ℃, stirs decolouring, and said extracting solution and activated carbon weight ratio are 300: 1～50: 1;
(4) extracting solution after step (3) processing is flow through cation-anion resin tandem exchange column to remove pigment and salt;
(5) with the extracting solution concentrating under reduced pressure after step (4) processing, the sugar weight percentage concentration is 30%～60% in the control liquid concentrator, in liquid concentrator, adds 95% ethanol of its 4～6 times of volumes again, and vibration shakes up 1～3h; Leave standstill 12～36h after the taking-up, obtain the trehalose white crystal.
Preferably, the weight percent concentration of alcohol is 30～50% in the said step (2), and temperature is 70～80 ℃, and extraction time is 60min～90min; Said yeast and alcohol weight ratio are 1: 20.
Preferably, the weight percent concentration of alcohol is 40% in the said step (2), and temperature is 75 ℃, and extraction time is 75min.
Preferably, pH is 4.0 in the said step (3), and temperature is 30 ℃, and churning time is 40min, and said extracting solution and activated carbon weight ratio are 100: 1.
Preferably, the sugar weight percentage concentration is 45% in the middle liquid concentrator of said step (5), and said alcoholic acid volume is 5 times of liquid concentrator, and vibration shakes up 2h, and time of repose is 24h.
Preferably, said Zeo-karb is 001x7, and anionite-exchange resin is 201x7.
Preferably, the said churning time of step (3) is 20～60min.
The present invention has following advantage and beneficial effect:
1, the existing technology of utilizing yeast production sterol is after having extracted sterol, and yeast is as offal treatment, and part factory directly is disposed in the water drain, causes the severe contamination of environment.The present invention reasonably utilizes discarded yeast to carry out deep processing, low in raw material cost, and help reducing pollution to environment.
2, be raw material with the discarded yeast behind the production sterol, therefrom extract trehalose, because yeast has carried out broken wall; Art breading such as enzyme deactivation; The present invention produces trehalose technology relatively with utilizing dry yeast, can reasonably be connected pre-treating technology, saves production cost.
Embodiment
Be described further in the face of the present invention down, the scope that the present invention requires to protect is not limited to the scope that embodiment explains.
Embodiment 1
Discarded yeast heating behind the production sterol is dried to powder again to remove residual solvent; Get 1Kg yeast and 10Kg weight percent concentration and be 20% alcohol and mix, being heated to temperature is 60 ℃, lixiviate 30min; The centrifugal yeast of removing of extracting solution adds the 200g activated carbon in above-mentioned extracting solution, be 3.5 at pH, and temperature is 25 ℃, and churning time is to decolour under the 20min condition; Extracting solution flows through sun-cloudy tandem ion exchange resin exchange column (post is 001x7 Zeo-karb 200mL+201x7 anionite-exchange resin 200mL) to remove pigment and salt; The effluent concentrating under reduced pressure, the sugar weight percentage concentration is 30% in the control liquid concentrator, in the 200mL liquid concentrator, adds 95% ethanol of 1200mL, vibration shakes up 1h.Take out the back and leave standstill 36h, obtain trehalose white crystal 58g in room temperature.
Embodiment 2
Discarded yeast heating behind the production sterol is dried to powder again to remove residual solvent; Get 1Kg yeast and 30Kg weight percent concentration and be 60% alcohol and mix, being heated to temperature is 80 ℃, lixiviate 120min; The centrifugal yeast of removing of extracting solution adds the 100g activated carbon in above-mentioned extracting solution, be 4.5 at pH, and temperature is 35 ℃, and churning time is to decolour under the 60min condition; Extracting solution flows through sun-cloudy tandem ion exchange resin exchange column (201x7 anionite-exchange resin 200mL+001x7 Zeo-karb 200mL) to remove pigment and salt; The effluent concentrating under reduced pressure, the sugar weight percentage concentration is 60% in the control liquid concentrator, in liquid concentrator (140mL), adds 95% ethanol of its 4 times of volumes, vibration shakes up 3h.Take out the back and leave standstill 12h, obtain trehalose white crystal 81g in room temperature.
Embodiment 3
Discarded yeast heating behind the production sterol is dried to powder again to remove residual solvent; Get 1Kg yeast and 15Kg weight percent concentration and be 50% alcohol and mix, being heated to temperature is 70 ℃, lixiviate 60min; The centrifugal yeast of removing of extracting solution adds the 100g activated carbon in above-mentioned extracting solution, be 3.8 at pH, and temperature is 28 ℃, and churning time is to decolour under the 30min condition; Extracting solution flows through sun-cloudy tandem ion exchange resin exchange column (post is 001x7 Zeo-karb 300mL+201x7 anionite-exchange resin 200mL) to remove pigment and salt; The effluent concentrating under reduced pressure, the sugar weight percentage concentration is 40% in the control liquid concentrator, in liquid concentrator (240mL), adds 95% ethanol of its 6 times of volumes, vibration shakes up 1.5h.Take out the back and leave standstill 18h, obtain trehalose white crystal 93g in room temperature.
Embodiment 4
Discarded yeast heating behind the production sterol is dried to powder again to remove residual solvent; Get 1Kg yeast and 25Kg weight percent concentration and be 30% alcohol and mix, being heated to temperature is 80 ℃, lixiviate 90min; The centrifugal yeast of removing of extracting solution adds the 200g activated carbon in above-mentioned extracting solution, be 4.2 at pH, and temperature is 32 ℃, and churning time is to decolour under the 50min condition; Extracting solution flows through sun-cloudy tandem ion exchange resin exchange column (post is 001x7 Zeo-karb 200mL+201x7 anionite-exchange resin 200mL+001x7 Zeo-karb 100mL) to remove pigment and salt; The effluent concentrating under reduced pressure, the sugar weight percentage concentration is 50% in the control liquid concentrator, in liquid concentrator (210mL), adds 95% ethanol of its 4 times of volumes, vibration shakes up 2.5h.Take out the back and leave standstill 30h, obtain trehalose white crystal 98g in room temperature.
Embodiment 5
Discarded yeast heating behind the production sterol is dried to powder again to remove residual solvent; Get 1Kg yeast and 20Kg weight percent concentration and be 40% alcohol and mix, being heated to temperature is 75 ℃, lixiviate 75min; The centrifugal yeast of removing of extracting solution adds the 100g activated carbon in above-mentioned extracting solution, be 4.0 at pH, and temperature is 30 ℃, and churning time is to decolour under the 40min condition; Extracting solution flows through sun-cloudy tandem ion exchange resin exchange column (post is 001x7 Zeo-karb 200mL+201x7 anionite-exchange resin 200mL+001x7 Zeo-karb 100mL) to remove pigment and salt; The effluent concentrating under reduced pressure, the sugar weight percentage concentration is 45% in the control liquid concentrator, in liquid concentrator (235mL), adds 95% ethanol of its 5 times of volumes, vibration shakes up 2h.Take out the back and leave standstill 24h, obtain trehalose white crystal 103g in room temperature.
Sugar weight percentage concentration detection method in the liquid concentrator:
Adopt Abbe refractometer assay method.
The trehalose method for detecting purity:
Instrument: high performance liquid chromatograph (being furnished with the differential detector); Moving phase de-gassing vessel and 0.45um millipore filtration; Chromatographic column: nh 2 column (4.6mm X 300mm, 5um); Analytical balance: sensibility reciprocal 0.0001g; Microsyringe: 10uL.
Reagent: redistilled water; Second eyeball (chromatographically pure); Trehalose standard substance (purity＞99.5000).
Step
1, standard substance preparation
Standard substance need in 60 ℃ of Constant Temp. Ovens, to be used for weighing behind the dry 5h.Take by weighing the trehalose standard substance) about 0.5g, be accurate to 0.0001g, with water dissolution and be settled to 50mL, shake up.Use the 0.45um filtering with microporous membrane, collect filtrating confession and measure usefulness.
2, specimen preparation
Sample needs in 60 ℃ of Constant Temp. Ovens, to be used for weighing behind the dry 5h.Take by weighing the about 0.5g of trehalose sample, be accurate to 0.0001g, with water dissolution and be settled to 50mL, shake up.Use the 0.45um filtering with microporous membrane, collect filtrating confession and measure usefulness.
3, the mensuration of sample
Moving phase is the second eyeball: water=70: 30.At the power supply of measuring of connecting the differential detector previous day, preheating is stable, loads onto chromatographic column, with the logical people's moving phase of flow velocity of 0.1mL/min, equilibrate overnight.Before formal sample introduction is analyzed, more than used moving phase input reference cell 20min, recover normal flow paths again, make moving phase, regulate flow velocity, walk baseline to 1.0mL/min through sample pool, treat baseline walk steady after, with standardized solution and the sample for preparing sample introduction 10uL respectively.According to the sugar component in the qualitative sample of the RT of standard substance,
Peak area per sample is with the percentage composition of external standard method calculating sugar component.
4, the result calculates
Content of trehalose calculates by following formula:
X＝(A
x·m
s/A
s·m
x)100
In the formula:
The X-content of trehalose, %;
A
x-sample peak area;
m
sOne standard substance quality, unit is gram (g);
A
s-standard substance peak area;
m
x-sample quality, unit is gram (g).
Calculation result keeps a decimal.
Table 1 trehalose yield and assay result
Learn through above-mentioned analyzing and testing: the trehalose yield for preparing according to method of the present invention is more than 5.8%, and content is more than 89%, and extract purity is high, explains that technology of the present invention has applications well and is worth.
Claims (7)
1. a method of from the discarded yeast behind the production sterol, extracting trehalose is characterized in that, comprises the steps:
(1) the discarded yeast heating behind the production sterol is dried to powder again to remove residual solvent;
(2) be that 20～60% alcohol mixes with yeast and weight percent concentration, be heated to 60～90 ℃, lixiviate 30min～120min; The centrifugal yeast of removing of extracting solution, described yeast and alcohol weight ratio are 1: (10～30);
(3) adding activated carbon in the extracting solution after step (2) is centrifugal, is 3.5～4.5 at pH, under the condition that temperature is 25～35 ℃, stirs decolouring, and said extracting solution and activated carbon weight ratio are 300: 1～50: 1;
(4) extracting solution after step (3) processing is flow through sun-the moon or male-female or sun-male-female ion exchange resin tandem exchange column to remove pigment and salt;
(5) with the extracting solution concentrating under reduced pressure after step (4) processing, the sugar weight percentage concentration is 30%～60% in the control liquid concentrator, in liquid concentrator, adds 95% ethanol of its 4～6 times of volumes again, and vibration shakes up 1～3h; Leave standstill 12～36h after the taking-up, obtain the trehalose white crystal.
2. method according to claim 1 is characterized in that, the weight percent concentration of alcohol is 30～50% in the said step (2), and temperature is 70～80 ℃, and extraction time is 60min～90min; Said yeast and alcohol weight ratio are 1: 20.
3. method according to claim 2 is characterized in that, the weight percent concentration of alcohol is 40% in the said step (2), and temperature is 75 ℃, and extraction time is 75min.
4. according to claim 1 or 2 or 3 described methods, it is characterized in that pH is 4.0 in the said step (3), temperature is 30 ℃, and churning time is 40min, and said extracting solution and activated carbon weight ratio are 100: 1.
5. method according to claim 4 is characterized in that, the sugar weight percentage concentration is 45% in the middle liquid concentrator of said step (5), and said alcoholic acid volume is 5 times of liquid concentrator, and vibration shakes up 2h, and time of repose is 24h.
6. method according to claim 5 is characterized in that, said Zeo-karb is 001x7, and anionite-exchange resin is 201x7.
7. method according to claim 6 is characterized in that, the said churning time of step (3) is 20～60min.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201110362849XA CN102504040A (en) | 2011-11-16 | 2011-11-16 | Method for extracting trehalose from waste yeast after sterol production |
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| CN201110362849XA CN102504040A (en) | 2011-11-16 | 2011-11-16 | Method for extracting trehalose from waste yeast after sterol production |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04360692A (en) * | 1991-06-07 | 1992-12-14 | Kanegafuchi Chem Ind Co Ltd | Production of trehalose |
| JPH0591890A (en) * | 1991-08-27 | 1993-04-16 | Kanji Matsumoto | Production of trehalose |
| WO1995009243A1 (en) * | 1993-09-28 | 1995-04-06 | Ústav Makromolekulární Chemie Akademie Ved C^¿Eské Republiky | METHOD OF PRODUCING α,α-TREHALOSE |
| CN1174200A (en) * | 1997-09-18 | 1998-02-25 | 大连理工大学 | Seaweed sugar producing process |
| CN1309131A (en) * | 2001-02-28 | 2001-08-22 | 中国科学院微生物研究所 | Process for preparing mycose from fermented waste |
-
2011
- 2011-11-16 CN CN201110362849XA patent/CN102504040A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04360692A (en) * | 1991-06-07 | 1992-12-14 | Kanegafuchi Chem Ind Co Ltd | Production of trehalose |
| JPH0591890A (en) * | 1991-08-27 | 1993-04-16 | Kanji Matsumoto | Production of trehalose |
| WO1995009243A1 (en) * | 1993-09-28 | 1995-04-06 | Ústav Makromolekulární Chemie Akademie Ved C^¿Eské Republiky | METHOD OF PRODUCING α,α-TREHALOSE |
| CN1174200A (en) * | 1997-09-18 | 1998-02-25 | 大连理工大学 | Seaweed sugar producing process |
| CN1309131A (en) * | 2001-02-28 | 2001-08-22 | 中国科学院微生物研究所 | Process for preparing mycose from fermented waste |
Non-Patent Citations (2)
| Title |
|---|
| 张雪莲: "从酵母中提取纯化海藻糖", 《中国优秀硕士学位论文全文数据库工程科技I辑》, 15 August 2005 (2005-08-15) * |
| 李静等: "从面包酵母中提取海藻糖的研究", 《食品与药品》, vol. 10, no. 3, 10 March 2008 (2008-03-10), pages 24 - 26 * |
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Application publication date: 20120620 |