CN102326796A - Method for producing oyster hydrolyzate through microbial fermentation - Google Patents
Method for producing oyster hydrolyzate through microbial fermentation Download PDFInfo
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- CN102326796A CN102326796A CN201110171792A CN201110171792A CN102326796A CN 102326796 A CN102326796 A CN 102326796A CN 201110171792 A CN201110171792 A CN 201110171792A CN 201110171792 A CN201110171792 A CN 201110171792A CN 102326796 A CN102326796 A CN 102326796A
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Abstract
The invention discloses a method for producing an oyster hydrolyzate through microbial fermentation, which is characterized in that a pacific oyster is fermented under the action of Aspergillus oryzae to prepare an oyster hydrolyzate. The preparation process comprises the following steps: taking fresh oyster meat, cleaning, homogenizing, and inoculating with the Aspergillus oryzae for fermentation; and after the fermentation is finished, carrying out filtration sterilization, and concentrating the filtrate to obtain the oyster hydrolyzate. By adopting a liquid fermentation mode, the method has the advantages of reasonable design of process parameters, high operability, low cost and high total nitrogen extraction rate and glycogen extraction rate, avoids the use of finished proteases and amylases, can make full use of oyster resources, and has better industrial application potential.
Description
Technical field
The present invention relates to the method that a kind of liquid fermentation oyster prepares the oyster hydrolyzate, belong to processing of aquatic products and field of microbial biotechnology.
Background technology
The oyster delicious flavour, nutritious, have the good reputation of " marine milk ", be that China classifies one of health care curative effect article of integration of drinking and medicinal herbs in the first batch as.Oyster albumen is higher up to 50%, 8 kind of contents of essential amino acids, accounts for 40% of total amino acid content, and required histidine and the arginine content of infant is higher, is a kind of good protein.Contain the taurine more than 3% in the what is more important oyster meat.Taurine has effects such as eliminating inflammation and expelling toxin, hepatic cholagogic, reducing blood lipid, promotion child brain development and Anshen Bunao.In addition, also contain in the oyster meat up to 22.41% have mineral matter and trace elements such as the active glycogen of important physiological, DHA (DHA) and eicosapentaenoic acid unrighted acids such as (EPA), vitamin and iron, zinc, calcium, manganese, selenium.
Oyster is one of China's four big cultivated shellfishes, propagates aboundresources in a large number artificially in the coastal area.At present, oyster processing and comprehensive utilization are also relatively backward, are main to eat and to process the biltong goods raw how, are processed into oyster sauce or other flavouring on a small quantity.The oyster deep processing prepares high value added product and rarely has report, and the phenomenon of " Higher output is not accompanied by a higher income " appears in therefore domestic oyster culture industry.Generation is easy to absorb behind the proteolysis, dissolubility is good, emulsibility is good, and has the polypeptide of multiple physiologically active, has improved the functional characteristic of protein, has improved the nutrition and the value of protein.
Pacific oyster is claimed long oyster, Japan real oyster again, wide warm euryhalinous inner bay kind.At present, China coastal provinces part is cultured this kind more.The enzymolysis property and the technology that comprise multiple oysters such as Pacific oyster, Crassostrea rivularis, ostrea talienwhanensis Crosse are studied both at home and abroad at present.With the oyster hydrolysis, can produce and have the active peptides that comprises antifatigue, multiple physiologically active such as anti-oxidant.The oyster hydrolysis that comprises of report all is directly to add the finished product protease hydrolytic at present.The finished product protease price is expensive, has increased production cost.
Summary of the invention
Technical problem to be solved by this invention is the deficiency to prior art, provide a kind of new technology more rationally, the method that effectively reduces cost, produce the oyster hydrolyzate applicable to the microbial fermentation of the big production of industry.
Technical problem to be solved by this invention is to realize through following technical scheme.The present invention is the method that a kind of microbial fermentation is produced the oyster hydrolyzate, is characterized in, it adopts aspergillus oryzae Aspergillus oryzae liquid fermentation method, and its concrete steps are following:
(1) preparation of oyster: buy fresh Pacific oyster, clean up, take out oyster meat, refiner homogenate;
(2) preparation fermentation substrate: it is 15% to the M/V of protein concentration that homogenate uses distilled water diluting, and with the NaOH adjusting pH value to 7.0 of 1M, 60 ℃ of pasteurization 30min get fermentation substrate;
(3) microbial fermentation: with the activation on the PDA slant medium of aspergillus oryzae Aspergillus oryzae bacterial classification, the picking spore inoculating is to the PDA fluid nutrient medium from the culture presevation inclined-plane after the activation, and 30 ℃, 120rpm are cultured to cell concentration and reach 10
7CFU/ml gets the fermentation strain seed liquor; By V/V 5% inoculum concentration the fermentation strain seed liquor is inoculated in fermentation substrate, liquid amount V/V 20%, rotating speed 140rpm, 30 ℃, fermentation 48h;
(4) separate: after the fermentation ends, remove thalline, not protein hydrolysate and other impurity with the milipore filter ultrafiltration of holding back 100KD, cleaner liquid is the oyster hydrolysate, perhaps the powder process of cleaner liquid concentrate drying is got powdery oyster hydrolysate.
Potato culture (PDA) can adopt following method configuration: potato 200g (section poach 30min crosses leaching juice), sucrose 20.0g, distilled water 1000mL, pH value nature.It is 2% agar that solid medium adds m/V in addition.
The aspergillus oryzae Aspergillus oryzae (bacterial strain AS3.951) that can select for use of the present invention available from Institute of Micro-biology of the Chinese Academy of Sciences.
Below be that the inventor did about the technical scheme of the inventive method or the optimization test and the result thereof of parameter.
1, the preparation of working sample:, boil the 10min enzyme that goes out respectively in the sampling of different fermentation time.Zymotic fluid is crossed 0.45 μ m miillpore filter, and filtrating is measured as sample.
2, inoculum concentration is to the influence of total nitrogen recovery rate and glycogen recovery rate:
After preparing fermentation substrate according to the inventive method step (2), inoculum concentration is respectively 2.5%, 5%, and 10%, 15%; Liquid amount is V/V 20%, rotating speed 120rpm, and 30 ℃ of fermented and cultured are respectively 0,12; 24,36,48,60; 72,84, the influence of inoculum concentration to total nitrogen recovery rate and glycogen recovery rate measured in the 96h sampling.Result such as Fig. 1, inoculum concentration variable effect total nitrogen recovery rate and glycogen recovery rate, inoculum concentration is 2.5% o'clock, the total nitrogen recovery rate the fermentation 60h reach maximum, and inoculum concentration be 5%, 10%, 15% o'clock all the fermentation 48h reach the higher extracted level; Inoculum concentration is 2.5% and 5% o'clock, and the glycogen recovery rate reaches the higher extracted level at fermentation 72h with 48h respectively, and inoculum concentration is 10% and 15% o'clock, and the two all reaches the higher extracted level about the 36h that ferments.Comprehensive total nitrogen recovery rate, glycogen recovery rate and cost, inoculum concentration selects 5%, and this condition bottom fermentation 48h total nitrogen recovery rate and glycogen recovery rate all reach the higher extracted level.
3, initial pH is to the influence of total nitrogen recovery rate and glycogen recovery rate:
With distilled water homogenate is diluted to protein concentration m/V 15%, is respectively 5.0,6.0,7.0,8.0,9.0,60 ℃, the 30min pasteurization with 1M NaOH or 1M salt acid for adjusting pH value.Inoculum concentration V/V 5%, liquid amount V/V 20%, and rotating speed 120rpm, 30 ℃ of fermented and cultured, respectively 0,12,24,36,48,60,72,84, the influence of initial pH to total nitrogen recovery rate and glycogen recovery rate measured in the 96h sampling.Result such as Fig. 2, the total nitrogen recovery rate reaches maximum the earliest when pH8, and glycogen recovery rate recovery rate when pH6 is the highest, and reach maximum the earliest, and balance is considered protein extracting ratio and glycogen recovery rate, selecting the fermentation initial pH value is 7.0.
4, concentration of substrate is to the influence of total nitrogen recovery rate and glycogen recovery rate:
With distilled water homogenate is diluted to protein concentration m/V 5%, 10%, 15%, 20%, regulates pH value to 7.0,60 ℃, 30min pasteurization with 1M NaOH.Inoculum concentration V/V5%, liquid amount V/V 20%, rotating speed 120rpm, 30 ℃ of fermented and cultured, respectively 0,12,24,36,48,60,72,84, the influence of initial protein concentration to total nitrogen recovery rate and glycogen recovery rate measured in the 96h sampling.Result such as Fig. 3, protein concentration be 5%, 10% and 15% o'clock not remarkable to the influence of total nitrogen recovery rate and glycogen recovery rate, all significantly descend but protein concentration is 20% o'clock total nitrogen recovery rate and glycogen recovery rate, therefore select protein concentration 15%.
5, fermentation temperature is to the influence of total nitrogen recovery rate and glycogen recovery rate:
With distilled water homogenate is diluted to protein concentration m/V 15%, regulates pH value to 7.0,60 ℃, 30min pasteurization with 1M NaOH.Inoculum concentration V/V 5%, liquid amount V/V 20%, and rotating speed 120rpm, respectively at 26 ℃, 28 ℃, 30 ℃, 32 ℃ of fermented and cultured, respectively 0,12,24,36,48,60,72,84, the influence of fermentation temperature to total nitrogen recovery rate and glycogen recovery rate measured in the 96h sampling.Result such as Fig. 4, temperature total nitrogen recovery rate and glycogen recovery rate in the time of 30 ℃ reach peak the earliest, and therefore selecting fermentation temperature is 30 ℃.
6, liquid amount is to the influence of total nitrogen recovery rate and glycogen recovery rate:
With distilled water homogenate is diluted to protein concentration m/V 15%, regulates pH value to 7.0,60 ℃, 30min pasteurization with 1M NaOH.Inoculum concentration V/V 5%, and liquid amount is adjusted into V/V 10%, 20%, 30%, 40% respectively, rotating speed 120rpm, and 30 ℃ of fermented and cultured are respectively 0; 12,24,36,48,60; 72,84, the influence of liquid amount to total nitrogen recovery rate and glycogen recovery rate measured in the 96h sampling.Result such as Fig. 5, liquid amount are that 20% o'clock total nitrogen recovery rate and glycogen recovery rate reach peak the earliest, and therefore selecting liquid amount is 20%.
7, rotating speed is to the influence of total nitrogen recovery rate and glycogen recovery rate:
With distilled water homogenate is diluted to protein concentration m/V 15%, regulates pH value to 7.0,60 ℃, 30min pasteurization with 1M NaOH.Inoculum concentration V/V 5%, and liquid amount V/V 20%, rotating speed are set at 100rpm, 120rpm, 140rpm, 160rpm respectively, and 30 ℃ of fermented and cultured are respectively 0; 12,24,36,48,60; 72,84, the influence of rotating speed to total nitrogen recovery rate and glycogen recovery rate measured in the 96h sampling.Result such as Fig. 6, total nitrogen recovery rate and glycogen recovery rate reached peak the earliest when rotating speed was 140rpm, and therefore selecting rotating speed is 140rpm.
8, fermentation time is to the influence of total nitrogen recovery rate, glycogen recovery rate, protease, amylase activity:
With distilled water homogenate is diluted to protein concentration m/V 15%, regulates pH value to 7.0,60 ℃, 30min pasteurization with 1M NaOH.Inoculum concentration V/V 5%, liquid amount V/V 20%, and rotating speed 140rpm, 30 ℃ of fermented and cultured, respectively 0,12,24,36,48,60,72,84, the influence of fermentation time to total nitrogen recovery rate, glycogen recovery rate, proteinase activity, amylase activity measured in the 96h sampling.Result such as Fig. 7, along with the prolongation of fermentation time, total nitrogen recovery rate, glycogen recovery rate, proteinase activity, amylase activity increase, and total nitrogen recovery rate, glycogen extraction rate reached are to maximum behind the fermentation 48h, and therefore selecting fermentation time is 48h.
9, hydrolyzate separates and measures: after the fermentation ends, can remove thalline and not protein hydrolysate and other impurity with the U.S. Millipore company milipore filter ultrafiltration of holding back 100KD.Measure the free amino acid and the taurine of ultrafiltrate and find that the ultrafiltrate free amino acid accounts for 35.91% of total amino acid, taurine accounts for 13.05% of total amino acid.
Compared with prior art, the inventive method adopts the liquid fermentation form, and its process parameters design is reasonable; Workable, need not to use finished product protease and amylase, cost is low; Total nitrogen recovery rate and glycogen recovery rate are high, can make full use of the oyster resource, have commercial Application potentiality preferably.
Description of drawings
Fig. 1 is the influence figure of inoculum concentration to total nitrogen recovery rate and glycogen recovery rate; Among the figure: ■; ●; ▲,
is respectively inoculum concentration is 2.5%, 5%; 10%, 15% o'clock total nitrogen recovery rate; ; Zero; △,
is respectively inoculum concentration is 2.5%, 5%; 10%, 15% o'clock glycogen recovery rate;
Fig. 2 is the influence figure of initial pH to total nitrogen recovery rate and glycogen recovery rate; Among the figure: ■; ●, ▲,
◆ being respectively initial pH is 5; 6; 7,8,9 o'clock total nitrogen recovery rates; , zero, △;
; It is 5,6,7 that ◇ is respectively initial pH; 8,9 o'clock glycogen recovery rates;
Fig. 3 is the influence figure of concentration of substrate to total nitrogen recovery rate and glycogen recovery rate; Among the figure: ■; ●; ▲,
is respectively concentration of substrate is 5%, 10%; 15%, 20% o'clock total nitrogen recovery rate; ; Zero; △,
is respectively concentration of substrate is 5%, 10%; 15%, 20% o'clock glycogen recovery rate; Fig. 4 is the influence figure of fermentation temperature to total nitrogen recovery rate and glycogen recovery rate; Among the figure: ■; ●; ▲,
is respectively fermentation temperature is 26 ℃, 28 ℃; 30 ℃, total nitrogen recovery rate in the time of 32 ℃; ; Zero; △,
is respectively fermentation temperature is 26 ℃, 28 ℃; 30 ℃, glycogen recovery rate in the time of 32 ℃
Fig. 5 is the influence figure of liquid amount to total nitrogen recovery rate and glycogen recovery rate; Among the figure: ■; ●; ▲,
is respectively liquid amount is 10%, 20%; 30%, 40% o'clock total nitrogen recovery rate; ; Zero; △,
is respectively liquid amount is 10%, 20%; 30%, 40% o'clock glycogen recovery rate;
Fig. 6 is the influence figure of rotating speed to total nitrogen recovery rate and glycogen recovery rate; Among the figure: ■; ●; ▲,
is respectively rotating speed is 100rpm, 120rpm; 140rpm, total nitrogen recovery rate during 160rpm; ; Zero; △,
is respectively rotating speed is 100rpm, 120rpm; 140rpm, glycogen recovery rate during 160rpm;
Fig. 7 is that fermentation time produces protease and the diastatic figure that influences to protein recovery, glycogen recovery rate and aspergillus oryzae; Among the figure: the ■ protein recovery; ● the glycogen recovery rate; ▲ proteinase activity;
amylase activity.
The specific embodiment
Below further describe concrete technical scheme of the present invention,, and do not constitute restriction its right so that those skilled in the art understands the present invention further.
Embodiment 1, and a kind of microbial fermentation prepares the method for oyster hydrolysate, and it adopts aspergillus oryzae Aspergillus oryzae liquid fermentation method, and its concrete steps are following:
(1) preparation of oyster: buy fresh Pacific oyster, clean up, take out oyster meat, refiner homogenate;
(2) preparation fermentation substrate: it is 15% to the M/V of protein concentration that homogenate uses distilled water diluting, and with the NaOH adjusting pH value to 7.0 of 1M, 60 ℃ of pasteurization 30min get fermentation substrate;
(3) microbial fermentation: with the activation on the PDA slant medium of aspergillus oryzae Aspergillus oryzae bacterial classification; The picking spore inoculating is to the PDA fluid nutrient medium from the culture presevation inclined-plane after the activation; 30 ℃, 120rpm are cultured to cell concentration and reach 107CFU/ml, the fermentation strain seed liquor; By V/V 5% inoculum concentration the fermentation strain seed liquor is inoculated in fermentation substrate, liquid amount V/V 20%, rotating speed 140rpm, 30 ℃, fermentation 48h; (4) separate: after the fermentation ends, remove thalline, not protein hydrolysate and other impurity with the milipore filter ultrafiltration of holding back 100KD, cleaner liquid is the oyster hydrolysate, perhaps the powder process of cleaner liquid concentrate drying is got powdery oyster hydrolysate.
Claims (1)
1. a microbial fermentation prepares the method for oyster hydrolysate, and it is characterized in that: it adopts aspergillus oryzae
Aspergillus oryzaeLiquid fermentation method, its concrete steps are following:
(1) preparation of oyster: buy fresh Pacific oyster, clean up, take out oyster meat, refiner homogenate;
(2) preparation fermentation substrate: it is 15% to the M/V of protein concentration that homogenate uses distilled water diluting, and with the NaOH adjusting pH value to 7.0 of 1M, 60 ℃ of pasteurization 30min get fermentation substrate;
(3) microbial fermentation: with aspergillus oryzae
Aspergillus oryzaeBacterial classification activation on the PDA slant medium, the picking spore inoculating is to the PDA fluid nutrient medium from the culture presevation inclined-plane after the activation, and 30 ℃, 120rpm are cultured to cell concentration and reach 10
7CFU/ml gets the fermentation strain seed liquor; By V/V 5% inoculum concentration the fermentation strain seed liquor is inoculated in fermentation substrate, liquid amount V/V 20%, rotating speed 140rpm, 30 ℃, fermentation 48h;
(4) separate: after the fermentation ends, remove thalline, not protein hydrolysate and other impurity with the milipore filter ultrafiltration of holding back 100KD, cleaner liquid is the oyster hydrolysate, perhaps the powder process of cleaner liquid concentrate drying is got powdery oyster hydrolysate.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102787079A (en) * | 2012-08-06 | 2012-11-21 | 江南大学 | Aspergillus oryzae and application method thereof in preparing sauce fragrant flavor material |
| CN104172080A (en) * | 2014-08-05 | 2014-12-03 | 北海富安源生物科技有限公司 | Method for extracting amino acid from oyster meat |
| CN105861565A (en) * | 2016-06-04 | 2016-08-17 | 中国海洋大学 | Turbot fishskin fermentation product with antioxidant activity |
| CN108396048A (en) * | 2018-02-08 | 2018-08-14 | 舟山海研食品科技有限公司 | The preparation method of oyster active peptides |
| CN109401983A (en) * | 2018-11-20 | 2019-03-01 | 佛山市海天(高明)调味食品有限公司 | A kind of aspergillus oryzae ZA151 and its application |
| JP2019218344A (en) * | 2018-06-15 | 2019-12-26 | チェジュ ナショナル ユニバーシティー インダストリー−アカデミック コーポレーション ファウンデーション | Composition for improving bone health, including functional fermented material using oysters |
| CN115777902A (en) * | 2022-12-07 | 2023-03-14 | 佛山市海天(宿迁)调味食品有限公司 | Composite-flavor oyster juice and oyster sauce and preparation method thereof |
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| JP2008142032A (en) * | 2006-12-12 | 2008-06-26 | Unitech Medical Co Ltd | Oyster extract and method of preparation of oyster extract |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102787079A (en) * | 2012-08-06 | 2012-11-21 | 江南大学 | Aspergillus oryzae and application method thereof in preparing sauce fragrant flavor material |
| CN102787079B (en) * | 2012-08-06 | 2014-09-17 | 江南大学 | Aspergillus oryzae and application method thereof in preparing sauce fragrant flavor material |
| CN104172080A (en) * | 2014-08-05 | 2014-12-03 | 北海富安源生物科技有限公司 | Method for extracting amino acid from oyster meat |
| CN104172080B (en) * | 2014-08-05 | 2015-11-18 | 北海富安源生物科技有限公司 | Amino acid whose method is extracted in oyster meat |
| CN105861565A (en) * | 2016-06-04 | 2016-08-17 | 中国海洋大学 | Turbot fishskin fermentation product with antioxidant activity |
| CN108396048A (en) * | 2018-02-08 | 2018-08-14 | 舟山海研食品科技有限公司 | The preparation method of oyster active peptides |
| JP2019218344A (en) * | 2018-06-15 | 2019-12-26 | チェジュ ナショナル ユニバーシティー インダストリー−アカデミック コーポレーション ファウンデーション | Composition for improving bone health, including functional fermented material using oysters |
| CN109401983A (en) * | 2018-11-20 | 2019-03-01 | 佛山市海天(高明)调味食品有限公司 | A kind of aspergillus oryzae ZA151 and its application |
| CN109401983B (en) * | 2018-11-20 | 2020-01-10 | 佛山市海天(高明)调味食品有限公司 | Aspergillus oryzae ZA151 and application thereof |
| CN115777902A (en) * | 2022-12-07 | 2023-03-14 | 佛山市海天(宿迁)调味食品有限公司 | Composite-flavor oyster juice and oyster sauce and preparation method thereof |
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