CN102326763B - Method for preparing yeast extract (YE) from Chinese red dates - Google Patents
Method for preparing yeast extract (YE) from Chinese red dates Download PDFInfo
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Abstract
The invention provides a method for preparing a yeast extract (YE) from Chinese red dates. The method comprises the following steps: preparing Chinese red date juice; preparing yeast seed liquid; preparing yeast sludge; breaking a yeast cell wall; carrying out enzyme inactivation; concentrating; and drying. Tests for the YE prepared by the embodiment 1 of the invention show that the YE yield is 9.78g/L, the dry matter content is 95.4%, the total nitrogen content is 5.12%, the amino acid nitrogen content is 2.39%, and the nucleic acid content is 0.41%. Therefore, the YE meets the requirements for flavor products shown in the standard GB/T 23530-2009 yeast extract and can be taken as a food seasoning.
Description
Technical field
The invention belongs to the food seasoning technical field, be specifically related to the preparation method of red date yeast extraction.
Background technology
Red date has another name called date, and is nutritious, with a long history, just classified as one of " five fruits " since ancient times.Red date meat multi-flavor is sweet, contains abundant nutrition such as carbohydrate, organic acid, vitamin, trace element, and wherein the most outstanding is that vitamin content is high, enjoys the good reputation of " natural complex ball ".Red date is not only nutritious, and has very high health-care medical value.Red date can improve monokaryon--the phagocytic function of phagocyte system in the body, protects liver in addition, strengthens the effect of muscle power, is one of fruit of liking very much of China people.Red date except that part is eaten raw, more in order to drying, so on the selling market, see dried jujube more.Owing to the economy of red date and the effect of ecological benefits, China's jujube cultivation area enlarges rapidly in recent years, and there is a sharp increase in output for red date; All there is feasible every year a large amount of red dates can not in time sell and remain; Cause huge waste, therefore, it is extremely urgent to solve red date deep processing problem.
Yeast extraction has another name called yeast extract, is to use the saccharomycete of food grade to be raw material, adopts biotechnology, after rich in protein, nucleic acid etc. are degraded in the yeast cells, and the natural flavouring of re-refining and processing.The world protein hydrolysate committee stipulated in 1977: " yeast extract is the food ingredient as natural flavouring, and its main component is the water-soluble substances of amino acid, peptide, polypeptide, flavour nucleotide and yeast cells ".Natural, nutrition, healthy yeast extraction are widely used in the various food as freshener and flavour enhanced dose, give food delicate flavour and mellow sense.At present, produce the raw material of yeast extraction or accessory substance saccharomyces cerevisiae that the prerequisite thing mainly contains Beer Production and the bread microzyme cultivated with blackstrap etc.Though lower with these raw material production yeast extraction costs, the preorder step is too much when producing yeast extraction, and is too loaded down with trivial details, is unfavorable for the large-scale production of yeast extraction.
Summary of the invention
Technical problem to be solved by this invention is to overcome the shortcoming of above-mentioned preparation yeast extraction method, provide a kind of prepared yeast extraction delicious, be of high nutritive value prepare the method for yeast extraction with red date.
Solving the problems of the technologies described above the technical scheme that is adopted comprises the steps:
1, preparation jujube juice
Extra dry red wine jujube or new fresh date are rejected the fruit that goes rotten, clean up, with pulverizer that red date is broken; Get broken red date, with the water of 6 times of red date quality, with lixiviate jar or jacketed pan 95～100 ℃ of heat treatments 1 hour; Isolate red date Normal juice with link-suspended basket centrifuge, in the jujube slag, add the water of 3 times of red date quality, pack into lixiviate jar or jacketed pan; 95～100 ℃ are incubated 30 minutes, isolate red date Normal juice with link-suspended basket centrifuge; The red date Normal juice that merges extracted twice; With centrifugal after the 200 purpose duplex strainer coarse filtration with the automatic discharging type centrifuge; Using the vacuum decompression concentration pan again is that to be evaporated to soluble solid under 0.08～0.09MPa condition be 15% in vacuum, is prepared into jujube juice.
2, preparation saccharomycete seed liquid
Saccharomyces cerevisiae or candida utili bacterium are transferred in the brewer's wort solid slant medium, cultivated 24～28 hours, and be transferred to after the cultivation and carry out enlarged culture in the triangular flask that brewer's wort is housed for 30 ℃; Cultivated 24～28 hours for 30 ℃; Nutrient solution in the triangular flask is transferred in the vial cultivates, the culture medium in the vial is that soluble solid is 10% jujube juice, 30 ℃ of enlarged culture 24～28 hours; After having cultivated the nutrient solution in the vial is transferred to enlarged culture in the seeding tank; Nutrient solution in the seeding tank is that soluble solid is 15% jujube juice, and inoculum concentration is that soluble solid is 2%～4%, 160～250 rev/mins of stirrings of 15% jujube juice volume; Cultivated 24～28 hours, and be prepared into saccharomycete seed liquid for 30 ℃.
Above-mentioned saccharomyces cerevisiae (Saccharomyces cerevisiae) and candida utili bacterium (Candida utilis) are that bacterial classification is preserved in Shaanxi Normal University food fermentation laboratory.
3, preparation saccharomycete mud
, the soluble solid of step 1 preparation adds NH in being 15% jujube juice
4H
2PO
4Or urea, silicone oil, every liter of soluble solid is to add NH in 15% the jujube juice
4H
2PO
4Or urea 1.98～7.72g, silicone oil 3g; Process the saccharomycete nutrient solution, with the saccharomycete nutrient solution with the tubular type steam sterilizer 125～130 ℃ of sterilizations 5 seconds, be cooled to 30 ℃; Be transported in the fermentation tank; Insert the saccharomycete seed liquid of saccharomycete nutrient solution volume 2%～4%, feed filtrated air, the maintenance ventilation rate is 1.0～1.4m
3/ (minm
3), cultivated 36～60 hours for 28～32 ℃, with the centrifugal zymotic fluid of disk centrifugal separator, add sterilized water in the deposition, centrifugal with disk centrifugal separator, obtain saccharomycete mud.
4, saccharomycete broken wall
The saccharomycete mud that step 3 obtains is transferred in the broken wall jar; Add the water of 2 times of saccharomycete shale amounts, process saccharomycete suspension, regulate the pH value to 7.0 of saccharomycete suspension with the NaOH aqueous solution of 0.1mol/L; Add the salt of saccharomycete suspension quality 3%; Stir, 50 ℃ of broken wall treatment 8 hours make saccharomycete self-dissolving broken wall.
5, enzymatic inactivation is handled
Saccharomycete suspension after step 4 broken wall treatment is placed heating tank, be warming up to 85～90 ℃, be incubated 15 minutes, centrifugal with disk centrifugal separator, obtain the saccharomycete hydrolyzate.
6, concentrate
Is that decompression concentrates under 0.08～0.09MPa condition with the saccharomycete hydrolyzate in vacuum, and the total nitrogen content that is concentrated into concentrate reaches till 5%, obtains the yeast extraction concentrate.
7, drying
In the yeast extraction concentrate, add the maltodextrin of its quality 20%, mix, carry out drying with centrifugal spray dryer, EAT is 110 ℃, is prepared into yeast extraction.
In the saccharomycete broken wall step 4 of the present invention; In order to improve the degradation rate of protein; With 50 ℃ of broken wall treatment of salt of saccharomycete suspension quality 3% after 8 hours; The papain that adds saccharomycete shale amount 0.5% again, 50 ℃ were continued broken wall treatment 24 hours, the enzyme work >=6000U of described papain.
In preparation saccharomycete seed liquid step 2 of the present invention, the nutrient solution in the vial is transferred to enlarged culture in the seeding tank that soluble solid is 15% jujube juice is housed, optimum inoculation amount is that soluble solid is 3% of 15% a jujube juice volume.
In preparation saccharomycete seed liquid step 2 of the present invention, the nutrient solution in the vial is transferred to is equipped with in the seeding tank that soluble solid is 15% jujube juice, best 190 rev/mins of stirrings were cultivated 24～28 hours, and were prepared into saccharomycete seed liquid for 30 ℃.
In preparation saccharomycete mud step 3 of the present invention, be the best NH of adding in 15% the jujube juice to the soluble solid of step 1 preparation
4H
2PO
4, silicone oil, every liter of soluble solid is the preferred NH of adding in 15% the jujube juice
4H
2PO
41.98g～7.72g, silicone oil 3g, every liter of soluble solid are the best NH of adding in 15% the jujube juice
4H
2PO
43.86g, silicone oil 3g.
In the preparation saccharomycete mud step 3 of the present invention, with the saccharomycete nutrient solution with the tubular type steam sterilizer 125～130 ℃ of sterilizations 5 seconds, be cooled to 30 ℃; Be transported in the fermentation tank; The preferred saccharomycete seed liquid that inserts saccharomycete nutrient solution volume 3% feeds filtrated air, and keeping ventilation rate the best is 1.2m
3/ (minm
3), cultivated 48 hours for 30 ℃.
The present invention is high with sugar content, have no side effect, aboundresources, cheap red date are raw material, through the culture yeasts bacterium, produces yeast extraction, enlarges the red date utilization rate, improves its economic worth.The yeast extraction that adopts the embodiment of the invention 1 preparation is through test; The yeast extraction yield is 9.78g/L, and dry matter content 95.4%, total nitrogen content are 9.92%; Amino-acid nitrogen is 3.96%; Nucleic acid content is 0.41%, has reached standard " GB/T 23530-2009 yeast extract " flavor type requirement of products, can be used as food seasoning.
The specific embodiment
To further explain of the present invention, but the invention is not restricted to these embodiment below in conjunction with specific embodiment.
Embodiment 1
1, preparation jujube juice
Extra dry red wine jujube or new fresh date are rejected the fruit that goes rotten, clean up, with pulverizer that red date is broken; Get broken red date, with the water of 6 times of red date quality, with lixiviate jar or jacketed pan 95～100 ℃ of heat treatments 1 hour; Isolate red date Normal juice with link-suspended basket centrifuge, in the jujube slag, add the water of 3 times of red date quality, pack into lixiviate jar or jacketed pan; 95～100 ℃ are incubated 30 minutes, isolate red date Normal juice with link-suspended basket centrifuge; The red date Normal juice that merges extracted twice; With centrifugal under 3000 rev/mins rotating speed after the 200 purpose duplex strainer coarse filtration with the automatic discharging type centrifuge; Obtain soluble solid and be 4%～5% red date Normal juice; Using the vacuum decompression concentration pan is that to be evaporated to soluble solid under 0.08～0.09MPa condition be 15% in vacuum, is prepared into jujube juice.
2, preparation saccharomycete seed liquid
Saccharomyces cerevisiae or candida utili bacterium are transferred in the brewer's wort solid slant medium, cultivated 24～28 hours, and be transferred to after the cultivation and carry out enlarged culture in the triangular flask that brewer's wort is housed for 30 ℃; Cultivated 24～28 hours for 30 ℃; Nutrient solution in the triangular flask is transferred in the 10L vial cultivates, the culture medium in the vial is that soluble solid is 10% jujube juice, 30 ℃ of enlarged culture 24～28 hours; After having cultivated the nutrient solution in the vial is transferred to enlarged culture in the 250L seeding tank of being with stirring; Nutrient solution in the seeding tank is that soluble solid is 15% jujube juice, and inoculum concentration is that soluble solid is 3%, 190 rev/min of stirring of 15% jujube juice volume; 30 ℃ of stir culture 24～28 hours are prepared into saccharomycete seed liquid.
Above-mentioned saccharomyces cerevisiae (Saccharomyces cerevisiae) and candida utili bacterium (Candida utilis) are that bacterial classification is preserved in Shaanxi Normal University food fermentation laboratory; Soluble solid is that 10% jujube juice is to be that 15% jujube juice thin up is processed by soluble solid in the step 1.
3, preparation saccharomycete mud
, the soluble solid of step 1 preparation adds NH in being 15% jujube juice
4H
2PO
4, silicone oil, every liter of soluble solid is to add NH in 15% the jujube juice
4H
2PO
43.86g, silicone oil 3g, process the saccharomycete nutrient solution, with the saccharomycete nutrient solution with the tubular type steam sterilizer 125～130 ℃ of sterilizations 5 seconds; Be cooled to 30 ℃; Be transported in the 5000L fermentation tank, the fermentation tank liquid amount is 4000L, inserts the saccharomycete seed liquid of saccharomycete nutrient solution volume 3%; Feed filtrated air, the maintenance ventilation rate is 1.0～1.4m
3/ (minm
3), 190 rev/mins of stirrings were cultivated 48 hours for 30 ℃; Using disk centrifugal separator is 5000 rev/mins of following centrifugal zymotic fluids 20 minutes at rotating speed, in deposition, adds the sterilized water of 10 times of its quality, carries out centrifugal clarification with disk centrifugal separator; Rotating speed is 5000 rev/mins; In deposition, add the sterilized water of 10 times of its quality again, use the disk centrifugal separator centrifugal clarification, obtain saccharomycete mud.
4, saccharomycete broken wall
The saccharomycete mud that step 3 obtains is transferred in the broken wall jar, added the water of 2 times of saccharomycete shale amounts, process saccharomycete suspension; With the pH value to 7.0 that the NaOH aqueous solution of 0.1mol/L is regulated saccharomycete suspension, add the salt of saccharomycete suspension quality 3% again, 60 rev/mins of stirrings; 50 ℃ of broken wall treatment 8 hours; Make saccharomycete self-dissolving broken wall, add the papain of saccharomycete shale amount 0.5% again, 50 ℃ were continued broken wall treatment 24 hours.
Enzyme work >=the 6000U of above-mentioned papain.
5, enzymatic inactivation is handled
Saccharomycete suspension after step 4 broken wall treatment is placed heating tank; Be warming up to 85～90 ℃; Be incubated 15 minutes; Enzyme in the saccharomycete hydrolyzate is carried out Passivation Treatment, used the saccharomycete suspension of disk centrifugal separator after rotating speed is that 6000 rev/mins centrifugal enzymatic inactivation is handled down 30 minutes, obtain the saccharomycete hydrolyzate.
6, concentrate
Is that decompression concentrates under 0.08～0.09MPa condition with the saccharomycete hydrolyzate in vacuum, and the total nitrogen content that is concentrated into concentrate reaches till 5%, gets the yeast extraction concentrate.
7, drying
In the yeast extraction concentrate, add the maltodextrin of its quality 20%, mix, carry out drying with centrifugal spray dryer, EAT is 110 ℃, is prepared into yeast extraction.
Prepared yeast extraction lighter color is yellow, and is Powdered, tests, and the test of various physical and chemical indexs is with reference to the GB/T23530-2009 assay method, and the test result of its physical and chemical index is seen table 1.
Table 1 yeast extraction physical and chemical index test result
Can know that by table 1 yeast extraction that adopts the embodiment of the invention 1 preparation has reached standard " GB/T23530-2009 yeast extract " flavor type requirement of products through test, can be used as food seasoning.
The sanitary index test is tested with reference to the QB2582-2003 assay method, and the test result of sanitary index is seen table 2.
Table 2 yeast extraction sanitary index test result
Because " GB/T 23530-2009 yeast extract " clearly do not stipulated the sanitary index of yeast extract, so this has been located with reference to " QB2582-2003 yeast extract ".Can know by table 2, adopt the yeast extraction of the embodiment of the invention 1 preparation, reach the requirement of " QB2582-2003 yeast extract ", can be used as food seasoning.
Nucleic acid determination is used ultraviolet absorption method, and the mensuration of aminoacid ingredient is utilized amino-acid analyzer, and various amino acid analysises are seen table 3.
Each seed amino acid percentage composition of table 3
Can be known that by table 3 yeast extraction that adopts the embodiment of the invention 1 preparation is through test, the amino acid kind is many, and content is abundant, is a kind of nutritious food seasoning.
Embodiment 2
In the preparation saccharomycete seed liquid step 2 of embodiment 1; Nutrient solution in the vial is transferred to enlarged culture in the 250L seeding tank of being with stirring; Nutrient solution in the seeding tank is that soluble solid is 15% jujube juice, and inoculum concentration is that soluble solid is 2%, 160 rev/min of stirring of 15% jujube juice volume; 30 ℃ of stir culture 24～28 hours, other steps of this step are identical with embodiment 1.Other steps are identical with embodiment 1, are prepared into yeast extraction.
Embodiment 3
In the preparation saccharomycete seed liquid step 2 of embodiment 1; Nutrient solution in the vial is transferred to enlarged culture in the 250L seeding tank of being with stirring; Nutrient solution in the seeding tank is that soluble solid is 15% jujube juice, and inoculum concentration is that soluble solid is 4%, 250 rev/min of stirring of 15% jujube juice volume; 30 ℃ of stir culture 24～28 hours, other steps of this step are identical with embodiment 1.Other steps are identical with embodiment 1, are prepared into yeast extraction.
Embodiment 4
In the preparation saccharomycete mud step 3 of embodiment 1～3,, the soluble solid of step 1 preparation adds NH in being 15% jujube juice
4H
2PO
4, silicone oil, every liter of soluble solid is to add NH in 15% the jujube juice
4H
2PO
41.98g, silicone oil 3g, process the saccharomycete nutrient solution, other steps of this step are identical with embodiment 1.Other steps are identical with corresponding embodiment, are prepared into yeast extraction.
Embodiment 5
In the preparation saccharomycete mud step 3 of embodiment 1～3,, the soluble solid of step 1 preparation adds NH in being 15% jujube juice
4H
2PO
4, silicone oil, every liter of soluble solid is to add NH in 15% the jujube juice
4H
2PO
47.72g, silicone oil 3g, process the saccharomycete nutrient solution, other steps of this step are identical with embodiment 1.Other steps are identical with corresponding embodiment, are prepared into yeast extraction.
Embodiment 6
In the preparation saccharomycete mud step 3 of embodiment 1～5; With the saccharomycete nutrient solution with the tubular type steam sterilizer 125～130 ℃ of sterilizations 5 seconds; Be cooled to 30 ℃; Be transported in the fermentation tank, insert the saccharomycete seed liquid of saccharomycete nutrient solution volume 2%, other steps of this step are identical with corresponding embodiment.Other steps are identical with corresponding embodiment, are prepared into yeast extraction.
Embodiment 7
In the preparation saccharomycete mud step 3 of embodiment 1～5; With the saccharomycete nutrient solution with the tubular type steam sterilizer 125～130 ℃ of sterilizations 5 seconds; Be cooled to 30 ℃; Be transported in the fermentation tank, insert the saccharomycete seed liquid of saccharomycete nutrient solution volume 4%, other steps of this step are identical with corresponding embodiment.Other steps are identical with corresponding embodiment, are prepared into yeast extraction.
Embodiment 8
In the preparation saccharomycete mud step 3 of embodiment 1～7, with the saccharomycete nutrient solution with the tubular type steam sterilizer 125～130 ℃ of sterilizations 5 seconds, be cooled to 30 ℃, be transported in the fermentation tank, feed filtrated air, the maintenance ventilation rate is 1.4m
3/ (minm
3), 250 rev/mins of stirrings were cultivated 36 hours for 32 ℃, and other steps of this step are identical with corresponding embodiment.Other steps are identical with corresponding embodiment, are prepared into yeast extraction.
Embodiment 9
In the preparation saccharomycete mud step 3 of embodiment 1～7, with the saccharomycete nutrient solution with the tubular type steam sterilizer 125～130 ℃ of sterilizations 5 seconds, be cooled to 30 ℃, be transported in the fermentation tank, feed filtrated air, the maintenance ventilation rate is 1.0m
3/ (minm
3), 160 rev/mins of stirrings were cultivated 60 hours for 28 ℃, and other steps of this step are identical with corresponding embodiment.Other steps are identical with corresponding embodiment, are prepared into yeast extraction.
Embodiment 10
In the preparation saccharomycete mud step 3 of embodiment 1～9, used NH
4H
2PO
4The urea replacement of quality such as use, other steps of this step are identical with corresponding embodiment.Other steps are identical with corresponding embodiment, are prepared into yeast extraction.
In order to confirm optimum process condition of the present invention, the inventor has carried out a large amount of laboratory research tests, and various test situation are following:
Experiment material: candida tropicalis bacterium (Candida tropicalis), buy in Shaanxi Institute of Microbiology; Candida utili bacterium (Candida utilis), saccharomyces cerevisiae (Saccharomyces cerevisiae) are preserved by Shaanxi Normal University food fermentation laboratory.
Laboratory apparatus: the LGJ-18C vacuum freeze drier, scientific instrument factory provides by Fourth Ring, Beijing; The Kjeltec2300 full-automatic Kjeldahl determination device is provided by Sweden FOX company; The full-automatic amino-acid analyzer of L-8900 is provided by Japanese HITACHI company; The DL-4C low speed large capacity centrifuge is provided by Anting Scientific Instrument Factory, Shanghai; The TU-1810 ultraviolet-uisible spectrophotometer is analysed general providing by Beijing is general; The RE-52 rotary evaporator is provided by last Hai'an booth laboratory apparatus Co., Ltd; SCP-9080MBE water isolation type constant incubator is provided by Medical Equipment Plant of Shanghai Boxun Industrial Co., Ltd.; HHW-21CU-600 electric heating constant temperature tank is provided by Shanghai Fuma Experiment Equipment Co., Ltd.; The accurate pH meter of PHS-3C is provided by Shanghai Precision Scientific Apparatus Co., Ltd; The THZ-C constant temperature oscillator, testing equipment factory provides by the Taicang, Jiangsu.
The measuring method:
(1) crude protein content is measured: Kjeldahl, operate according to GB/T5009.5-2003.
(2) amino acid nitrogen content is measured: formol titration
Sample is used dissolved in distilled water; Draw sample solution 5mL; Add 55mL water, under agitation using NaOH aqueous solution volumetric soiutions to the pH value of 0.05mol/L is 8.20, adds formalin 10mL; Using NaOH aqueous solution volumetric soiutions to the pH value of 0.05mol/L is 9.20, and record consumes the volume V of the NaOH aqueous solution
3Do blank test with distilled water simultaneously, record adds the volume V of the 0.05mol/L NaOH aqueous solution that is consumed after the formalin
4
X in the formula
3Amino acid nitrogen content for sample; C
2Be the concentration of the NaOH aqueous solution, the mol/L of unit; V
3After adding formalin, the titration sample consumes the volume of the NaOH aqueous solution, Unit; V
4After adding formalin, blank test consumes the volume of the NaOH aqueous solution, Unit; N is the sample extension rate; M is the quality of sample, the g of unit; X
1Dry matter content for sample.
(3) aminoacid ingredient is measured: adopt amino-acid analyzer, pillar is 4.6 * 60mm (filler 2622) cationic ion-exchange resin packed column, the splitter temperature: 57 ℃, and the reaction column temperature: 135 ℃, column pressure: 4.4MPa, N
2Pressure: 0.11MPa, the buffer solution flow velocity: 0.4 μ L/ minute, the ninhydrin solution flow velocity: 0.35 μ L/ minute, buffer solution changed 7 times, sample size 20 μ L, 53 minutes analysis times.
(4) nucleic acid content is measured: ultraviolet absorption method
Determinand is diluted to suitable concentration, measures its ultraviolet absorption value A at the 260nm place
260, be calculated as follows the nucleic acid content of determinand:
0.024 is that concentration is the OD value of the nucleic acid solution of 1 μ g/mL in the formula.
1, bacterial classification enlarged culture
Saccharomyces cerevisiae, candida utili bacterium and candida tropicalis bacterium are transferred to respectively in the brewer's wort solid slant medium; Cultivated 24～28 hours for 30 ℃; After the inclined-plane is cultivated and is finished; Be transferred to respectively and carry out enlarged culture in the triangular flask that malt juice liquid medium is housed, saccharomyces cerevisiae and candida tropicalis bacterium be 30 ℃ of enlarged culture 24～28 hours, and the candida utili bacterium was 25 ℃ of enlarged culture 24～28 hours; Be transferred to respectively again after in malt juice liquid medium, cultivate finishing enlarged culture in the triangular flask that soluble solid is 10% jujube juice is housed, make saccharomycete seed liquid.
2, the optimization of saccharomycete condition of culture
(1) comparison of different yeast strains growth result in jujube juice
With the 50mL soluble solid is that 10% jujube juice is packed in the 250mL triangular flask; PH keeps nature, inserts the saccharomycete seed liquid that makes in the test 1 of jujube juice volume 3% respectively, uses the shaking table shaken cultivation; The cultivation temperature of candida utili bacterium is 25 ℃; The cultivation temperature of saccharomyces cerevisiae and candida tropicalis bacterium is 30 ℃, and rotating speed is 160 rev/mins, shaken cultivation 50 hours.Use disk centrifugal separator to be 5000 rev/mins and descended centrifugal 20 minutes, after the deposition vacuum freeze drying, measure its crude protein content, relatively the growing state of 3 kinds of bacterial strains in jujube juice at rotating speed.Result of the test is seen table 4.
The comparison of the different yeast strains of table 4 growth result in jujube juice
Can be found out that by table 4 when saccharomyces cerevisiae and candida utili bacterium were culture medium with the jujube juice, albuminiferous ability was higher than the candida tropicalis bacterium far away, the white ability of laying eggs of saccharomyces cerevisiae is up to 2.938g/L.Therefore, the present invention selects for use saccharomyces cerevisiae and candida utili bacterium as the bacterial strain of producing yeast extraction.
(2) jujube juice concentration is to cultivating the influence of brewer's yeast bacteria growing
The 50mL jujube juice is packed in the 250mL triangular flask, and pH keeps nature, and adjusting its soluble solid respectively is 5%, 10%, 15%, 20%, 25%, inserts the saccharomyces cerevisiae seed liquor after test 1 enlarged culture, and inoculum concentration is 3% of a jujube juice volume.Other experimental conditions are identical with test (1), relatively the growing state of saccharomyces cerevisiae in the jujube juice of different soluble solids.Result of the test is seen table 5.
Table 5 jujube juice concentration is to cultivating the influence of brewer's yeast bacteria growing
Can know by table 5; When soluble solid content when 5% is increased to 20%; Crude protein content increases and increases along with soluble solid content, still, and when soluble solid content is increased to 25%; The crude protein content of thalline descends on the contrary, and the jujube juice that high concentration is described is unfavorable for the growth of thalline because of osmotic pressure is too high.Soluble solid content is 15%～20% o'clock in the jujube juice, and the crude protein content of thalline is very nearly the same.Therefore, to select soluble solid be 15% the jujube juice nutrient solution as saccharomyces cerevisiae in the present invention.
(3) cultivation temperature is to the influence of saccharomyces cerevisiae
With the 50mL soluble solid is that 15% jujube juice is packed in the 250mL triangular flask; PH keeps nature; Saccharomyces cerevisiae seed liquor after access test 1 enlarged culture, inoculum concentration is 3% of a jujube juice volume, respectively 26 ℃, 28 ℃, 30 ℃, 32 ℃, 34 ℃ enlarged culture.Other experimental conditions are identical with test (1), relatively the growing state of saccharomyces cerevisiae under different cultivation temperature.
Result of the test is seen table 6.
Table 6 cultivation temperature is to the influence of saccharomyces cerevisiae
Can find out from table 6; When cultivation temperature is lower than 30 ℃; The crude protein content of thalline raises along with the rising of temperature, and when temperature was higher than 30 ℃, the crude protein content of thalline was along with the rising of temperature reduces on the contrary; Explain too high cultivation temperature to make endobacillary enzymatic inactivation and be suppressed, cause crude protein content to reduce.Therefore, to select the cultivation temperature of saccharomyces cerevisiae be 30 ℃ in the present invention.
(4) jujube juice pH is to cultivating the influence of saccharomyces cerevisiae
With the 50mL soluble solid is that 15% jujube juice is packed in the 250mL triangular flask; Using its pH value of citric acid and NaOH adjustment respectively is 4.0,4.5,5.0,5.5,6.0; Saccharomyces cerevisiae seed liquor after access test 1 enlarged culture, inoculum concentration is 3% of a jujube juice volume.Other experimental conditions are identical with test (1), relatively the growing state of saccharomyces cerevisiae in the jujube juice of different pH.
Result of the test is seen table 7.
Table 7 jujube juice pH is to cultivating the influence of saccharomyces cerevisiae
Can learn that from table 7 when the pH of jujube juice was between 4.0～5.5, the crude protein content of thalline increased along with the rising of pH, when pH was higher than 5.0, the crude protein content of thalline descended on the contrary.Soluble solid is that the optimal pH of 15% jujube juice culture yeasts bacterium is 4.5, and the saccharomycetic crude protein content of cultivation reaches 3.307g/L.Generally speaking, soluble solid is that the pH of 15% jujube juice is 4.5～5.0.Therefore, the present invention only need keep the natural pH of jujube juice, need not to adjust again, can satisfy the requirement of saccharomycete growth.
(5) inoculum concentration is to the influence of saccharomyces cerevisiae growth result
With the 50mL soluble solid is that 15% jujube juice is packed in the 250mL triangular flask, and jujube juice pH keeps nature, inserts the saccharomyces cerevisiae seed liquor after test 1 enlarged culture, and inoculum concentration is respectively 1%, 2%, 3%, 4%, 5% of jujube juice volume.Other experimental conditions are identical with test (1), relatively the growing state of saccharomyces cerevisiae in jujube juice of different vaccination amount.Result of the test is seen table 8.
Table 8 inoculum concentration is to the influence of saccharomyces cerevisiae growth result
Can be known that by table 8 crude protein content of thalline increases along with the increase of inoculum concentration, still, after inoculum concentration reached 3%, crude protein content increased not obvious.Therefore, consider that from the angle of economy it is 2%～4% of red date volume that the present invention selects the inoculum concentration of saccharomyces cerevisiae, the best is 3%.
(6) shaking speed is to the influence of brewer's yeast bacteria growing
With the 50mL soluble solid is that 15% jujube juice is packed in the 250mL triangular flask; The pH of jujube juice keeps nature, the saccharomyces cerevisiae seed liquor after access test 1 enlarged culture, and inoculum concentration is 3% of a jujube juice volume; The rotating speed that shaking table is set respectively is 130,160,190,220,250 rev/mins; Other experimental conditions are identical with test (1), carry out the shaken cultivation saccharomycete, and more different shaking speed are to the influence of brewer's yeast bacteria growing.Result of the test is seen table 9.
Table 9 shaking speed is to the influence of saccharomyces cerevisiae growth result
Can know that by table 9 along with the increase of shaking speed, the crude protein content of thalline increases thereupon; When shaking speed increases to 190 rev/mins; The crude protein content of thalline increases not obvious, and therefore, it is 160～250 rev/mins that the present invention selects shaking speed; The best is 190 rev/mins, and the ventilation rate of when rotating speed is 190 rev/mins, measuring in the nutrient solution is 1.2m
3/ (minm
3).
(7) incubation time is to the influence of brewer's yeast bacteria growing
With the 50mL soluble solid is that 15% jujube juice is packed in the 250mL triangular flask; The pH of jujube juice keeps nature; Saccharomyces cerevisiae seed liquor after access test 1 enlarged culture, inoculum concentration is 3% of a jujube juice volume, shaken cultivation is 24,36,48,60,72 hours respectively; Other experimental conditions are identical with test (1), and relatively incubation time is to the influence of saccharomyces cerevisiae growth result.Result of the test is seen table 10.
Table 10 incubation time is to the influence of saccharomyces cerevisiae growth result
Can know that by table 10 along with the prolongation of incubation time, the crude protein content of thalline increases thereupon.After incubation time extended to 48 hours, the crude protein content of thalline increased not obvious.Therefore, it is 15% jujube juice that the present invention uses soluble solid, and through the shaken cultivation saccharomyces cerevisiae, incubation time is 36～60 hours, and the best is 48 hours.
(8) nitrogenous source of nutrient solution is to the influence of brewer's yeast bacteria growing
With the 50mL soluble solid is that 15% jujube juice is packed in the 250mL triangular flask, and the pH of jujube juice keeps nature, inserts the saccharomyces cerevisiae seed liquor after test 1 enlarged culture, and inoculum concentration is 3% of a jujube juice volume, in nutrient solution, adds urea, KNO respectively
3, NH
4H
2PO
4, NH
4NO
3Four kinds of nitrogen sources, the addition of four kinds of nitrogenous sources all calculate KNO by the nitrogen content that every liter of nutrient solution contains 1g urea
3Addition is 3.36g/L, NH
4H
2PO
4Be 3.86g/L, NH
4NO
3Be 1.343g/L, other experimental conditions are identical with test (1), and relatively nitrogenous source is to the influence of saccharomyces cerevisiae growth result.Result of the test is seen table 11.
Table 11 nitrogenous source is to the influence of saccharomyces cerevisiae growth result
Can know by table 11, after adding different nitrogenous sources in the nutrient solution, through after the cultivation of certain hour; The thalline crude protein content of collecting all improves accordingly; When explaining, add the saccharomycetic growth and breeding of promotion that nitrogen source can be in various degree, improve its output with jujube juice culture yeasts bacterium.From table, can know, can improve the thalline crude protein content largely behind the interpolation urea, add NH
4H
2PO
4Back thalline crude protein content is the highest, reaches 4.560g/L, therefore, when the present invention uses jujube juice as nutrient solution culture yeasts bacterium, can select urea or NH
4H
2PO
4As the additional nitrogenous source of nutrient solution, optimal selection is NH
4H
2PO
4
(9) NH
4H
2PO
4Confirming of addition
With soluble solid is that 15% jujube juice is that nutrient solution is cultivated saccharomyces cerevisiae, respectively by add 0 in every liter of nutrient solution, 1.98g, 3.86g, 7.72g NH
4H
2PO
4Make an experiment, other experimental conditions are identical with test (1).Research variable concentrations NH
4H
2PO
4To cultivating the influence of saccharomyces cerevisiae, result of the test is seen table 12.
Table 12 NH
4H
2PO
4Confirming of addition
Can know by table 12, when in every liter of nutrient solution, adding 1.98g NH
4H
2PO
4The time, the crude protein content of saccharomyces cerevisiae and in nutrient solution, add nitrogenous source and be more or less the same is when in every liter of nutrient solution, adding NH
4H
2PO
4Amount during greater than 3.86g, the crude protein content of saccharomyces cerevisiae increases not obvious.Therefore, to select every liter of soluble solid be to add NH in 15% the jujube juice in the present invention
4H
2PO
41.98～7.72g, the best is 3.86g.
2, confirm the wall-breaking method of yeast cell
The saccharomycete flavor substance is present in the cell, discharges and extracts its flavor substance, must be with the cell membrane hydrolysis, break.Because the yeast cell wall is thicker, be difficult to break, must make it to destroy or decompose with external force.This test is with reference to microorganism wall-breaking method commonly used; Select for use autolysis method, ultrasonic method, multigelation method and four kinds of methods of alkaline process to make an experiment; Relatively four kinds of methods confirm from cultivate the saccharomycete that obtains, to extract the suitable wall-breaking method of flavor substance to saccharomycetic shell-broken effect.The concrete test operation step of four kinds of wall-breaking methods is following:
(1) autolysis method: the sterilized water that in the saccharomyces cerevisiae mud of centrifugal collection, adds 2 times of its quality; Process saccharomycete suspension; 0.1mol/L the NaOH aqueous solution pH value to 7.0 of regulating saccharomycete suspension, the salt of interpolation saccharomycete suspension quality 3% is incubated 50 ℃; With shaking table oscillation treatment 48 hours, shaking speed was 60 rev/mins.
(2) ultrasonic method: in the saccharomyces cerevisiae mud of centrifugal collection, add the sterilized water of 0.15 times of its quality, process saccharomycete suspension, be incubated 40 ℃, supersonic frequency is 40KHz, sonicated 1 hour.
(3) multigelation method: in the saccharomyces cerevisiae mud of centrifugal collection, add the sterilized water of 0.2 times of its quality, multigelation 3 times, each freeze-off time 2 hours, thawing time 30 minutes.
(4) alkaline process: in the saccharomyces cerevisiae mud of centrifugal collection, add the sterilized water of 0.12 times of its quality, process saccharomycete suspension, add the NaOH of saccharomycete suspension quality 3.5%, handled 2 hours for 100 ℃.
Processing is settled to equal volume with four kinds of treatment fluids after finishing, and measures the crude protein content in the treatment fluid, and result of the test sees Table 13.
The different saccharomycete wall-breaking methods of table 13 are to the influence of saccharomycete shell-broken effect
Can find out by table 13; Saccharomyces cerevisiae mud after the cultivation carries out the effect of broken wall treatment apparently higher than ultrasonic method and multigelation method according to autolysis method and alkaline process; The protein content of stripping is higher, alkaline process broken wall particularly, and the crude protein content in the saccharomycete hydrolyzate of broken wall reaches 4.501g/L.But the yeast extraction of the present invention's preparation is mainly used in food dressing, and after handling with alkaline process, not only the alkali flavor is obvious for the saccharomycete hydrolyzate, and serious to the destruction of the nutritional labeling in the hydrolyzate, and therefore, the alkaline process wall-breaking method also is not suitable for the present invention.Take all factors into consideration, the selected autolysis method of the present invention is to the saccharomycete broken wall.In order to improve the degradation rate of protein, can in the self-dissolving process, add papain, the auxiliary intracellular protease acting in conjunction of saccharomyces cerevisiae degrade proteins.Therefore, saccharomycete self-dissolving broken wall treatment is after 8 hours, add again saccharomycete shale amount 0.5% papain (enzyme lives >=6000U), 50 ℃ were continued broken wall treatment 24 hours, the saccharomycete hydrolyzate.
Claims (7)
1. one kind prepares the method for yeast extraction with red date, it is characterized in that it comprises the steps:
(1) preparation jujube juice
Extra dry red wine jujube or new fresh date are rejected the fruit that goes rotten, clean up, with pulverizer that red date is broken; Get broken red date, add the water of 6 times of red date quality, with lixiviate jar or jacketed pan 95～100 ℃ of heat treatments 1 hour; Separate red date Normal juice, in the jujube slag, add the water of 3 times of red date quality, pack into lixiviate jar or jacketed pan; 95～100 ℃ of heat treatment 30 minutes separates red date Normal juice, merges the red date Normal juice of extracted twice; It is concentrated to filter, centrifugalize, reduce pressure, and being concentrated into soluble solid is 15%, is prepared into jujube juice;
(2) preparation saccharomycete seed liquid
Saccharomyces cerevisiae or candida utili bacterium are transferred in the brewer's wort solid slant medium; Cultivated 24～28 hours for 30 ℃; Be transferred to after the cultivation in the triangular flask that brewer's wort is housed, 30 ℃ of enlarged culture 24～28 hours are transferred to the nutrient solution in the triangular flask and are equipped with in the vial that soluble solid is 10% jujube juice; 30 ℃ of enlarged culture 24～28 hours; The nutrient solution in the vial is transferred to again enlarged culture in the seeding tank that soluble solid is 15% jujube juice is housed, inoculum concentration is that soluble solid is 2%～4%, 160～250 rev/mins of stirrings of 15% jujube juice volume; Cultivated 24～28 hours, and be prepared into saccharomycete seed liquid for 30 ℃;
(3) preparation saccharomycete mud
, the soluble solid of step (1) preparation adds NH in being 15% jujube juice
4H
2PO
4Or urea, silicone oil, every liter of soluble solid is to add NH in 15% the jujube juice
4H
2PO
4Or urea 1.98～7.72g, silicone oil 3g, process the saccharomycete nutrient solution; With the saccharomycete nutrient solution with the tubular type steam sterilizer 125～130 ℃ of sterilizations 5 seconds, be cooled to 30 ℃, be transported in the fermentation tank, insert the saccharomycete seed liquid of saccharomycete nutrient solution volume 2%～4%, feed filtrated air, the maintenance ventilation rate is 1.0～1.4m
3/ (minm
3), 160～250 rev/mins of stirrings were cultivated 36～60 hours for 28～32 ℃, and the centrifugation zymotic fluid adds sterilized water in the deposition, centrifugal, obtains saccharomycete mud;
(4) saccharomycete broken wall
The saccharomycete mud that step (3) obtains is transferred in the broken wall jar; Add the water of 2 times of saccharomycete shale amounts; Process saccharomycete suspension,, add the salt of saccharomycete suspension quality 3% with the pH value to 7.0 that the NaOH aqueous solution of 0.1mol/L is regulated saccharomycete suspension; Stir 50 ℃ of broken wall treatment 8 hours;
(5) enzymatic inactivation is handled
Saccharomycete suspension after step (4) broken wall treatment is placed heating tank, be warming up to 85～90 ℃, be incubated 15 minutes, centrifugation obtains the saccharomycete hydrolyzate;
(6) concentrate
The saccharomycete hydrolyzate decompression that step (5) is obtained concentrates, and the total nitrogen content that is concentrated into concentrate is 5%, obtains the yeast extraction concentrate;
(7) drying
In the yeast extraction concentrate, add the maltodextrin of its quality 20%, mix, carry out drying with centrifugal spray dryer, EAT is 110 ℃, is prepared into yeast extraction.
2. the method for preparing yeast extraction with red date according to claim 1; It is characterized in that: in saccharomycete broken wall step (4); Add the salt of saccharomycete suspension quality 3%, stir, 50 ℃ of broken wall treatment 8 hours; The papain that adds saccharomycete shale amount 0.5% again, 50 ℃ were continued broken wall treatment 24 hours.
3. the method for preparing yeast extraction with red date according to claim 1; It is characterized in that: in preparation saccharomycete seed liquid step (2); Nutrient solution in the vial is transferred to enlarged culture in the seeding tank that soluble solid is 15% jujube juice is housed, inoculum concentration is that soluble solid is 3% of 15% a jujube juice volume.
4. according to claim 1 or the 3 described methods that prepare yeast extraction with red date; It is characterized in that: in preparation saccharomycete seed liquid step (2); Nutrient solution in the vial is transferred to 190 rev/mins of stirrings are housed in the seeding tank that soluble solid is 15% jujube juice.
5. according to claim 1ly prepare the method for yeast extraction, it is characterized in that: in preparation saccharomycete mud step (3), in the soluble solid of step (1) preparation is 15% jujube juice, add NH with red date
4H
2PO
4, silicone oil, every liter of soluble solid is to add NH in 15% the jujube juice
4H
2PO
41.98g～7.72g, silicone oil 3g.
6. according to claim 1ly prepare the method for yeast extraction, it is characterized in that: in preparation saccharomycete mud step (3), in the soluble solid of step (1) preparation is 15% jujube juice, add NH with red date
4H
2PO
4, silicone oil, every liter of soluble solid is to add NH in 15% the jujube juice
4H
2PO
43.86g, silicone oil 3g.
7. the method for preparing yeast extraction with red date according to claim 1; It is characterized in that: in preparation saccharomycete mud step (3), with the saccharomycete nutrient solution with the tubular type steam sterilizer 125～130 ℃ of sterilizations 5 seconds, be cooled to 30 ℃; Be transported in the fermentation tank; Insert the saccharomycete seed liquid of saccharomycete nutrient solution volume 3%, feed filtrated air, the maintenance ventilation rate is 1.2m
3/ (minm
3), 190 rev/mins of stirrings were cultivated 48 hours for 30 ℃.
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