CN105532855A - Novel polypeptide preservative and preparation method thereof - Google Patents
Novel polypeptide preservative and preparation method thereof Download PDFInfo
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- CN105532855A CN105532855A CN201510937458.4A CN201510937458A CN105532855A CN 105532855 A CN105532855 A CN 105532855A CN 201510937458 A CN201510937458 A CN 201510937458A CN 105532855 A CN105532855 A CN 105532855A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B7/00—Preservation or chemical ripening of fruit or vegetables
- A23B7/14—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10
- A23B7/153—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of liquids or solids
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Abstract
The present invention provides a novel polypeptide preservative and a preparation method thereof. The preparation method comprises the following steps: Tibet Kefir grains at the late logarithmic growth stage are inoculated into pure milk, the pure milk is subjected to sealed static fermentation at 30-40 DEG C until a part of curd is produced; the pure milk is added, a liquid fermented milk is drawn in a sterile environment and coated on a MC-solid medium and a MRS solid medium plate respectively, and the mixture is then sealed and cultured; after completion of the culture, bacterial colonies and the medium in the culture dish are added into pure milk, and at the same time, carbon and nitrogen sources are added, and the mixture is placed in a temperature of 30-35 DEG C to be fermented for 65-75 hours; and after the completion of the fermentation, the fermented milk is centrifuged to remove precipitates, the pH value of the resulted supernatant is adjusted to 2-5, the treated fermented milk is maintained at 8-10 DEG C for 60-120 min, then the fermented milk is centrifuged, the obtained precipitate is dried into a powder, and the powder is subjected to cryopreservation, thereby obtaining the natural novel polypeptide preservative. The present invention also provides a use method of the natural novel polypeptide preservative. The novel polypeptide preservative is safe and hygienic, natural and non-toxic, high-efficient, and broad in spectrum and stable in performance, and can be widely applied in the field of fresh fruit preservative.
Description
Technical field
The present invention relates to a kind of preservative technology field, be specifically related to a kind of novel polypeptide anticorrisive agent and preparation method thereof and its new opplication at fruit antisepsis fresh-keeping aspect.
Background technology
China is Production of fruit big country in the world, and the kind of current China fruit and the equal world rankings the first of output, therefore, Production of fruit is the mainstay industry of Chinese national economy, is the staple product of substantial connection people's lives and foreign exchange earning.But, if fruit is without suitable Preservation Treatment, problems such as there will be the decline of quality and local flavor in the processes such as storage, transport, go mouldy, rot, thus cause great economic loss.China has the fruit of 8,000 ten thousand tons to rot every year, loss nearly 80,000,000,000 yuan of total value.
Along with the development of science and technology, the raising of people's living standard, people pay attention to freshness and the nutritive value of fruit more, more and more higher requirement is proposed to the edible quality of fruit, thus also just more and more higher to the requirement of fruit freshness preserving technology, also pay attention to the use of antiseptic and fresh-keeping agent for fruits gradually simultaneously.At present, in production practices, mainly use chemical preservation antistaling agent, but frequently or excessively use chemical preservation antistaling agent can cause the problems such as microorganism resistance, chemical preservation antistaling agent are residual, fruit quality change, there is food security hidden danger, the requirement of people to High Quality Fruit can not be met, therefore, the development and application of the crude antistaling agent of safety economy, wide spectrum, efficient, stable performance has caused the concern of people, becomes a focus of food scientific research and application.
Summary of the invention
The object of the present invention is to provide a kind of safety and sanitation, Nantural non-toxic, high-efficiency broad spectrum, the novel polypeptide antisepsis antistaling agent of stable performance, another object is to provide the extensive use of this antisepsis antistaling agent in the fresh-keeping field of fruit antisepsis.This novel polypeptide antisepsis antistaling agent take plain chocolate as raw material, and the Tibet Kefir granule added after process ferments, then a kind of novel polypeptide antisepsis antistaling agent of obtaining of separation and purification.It has good inhibition to fruit spoilage organisms such as staphylococcus aureus, salmonella, Escherichia coli with to fruit postharvest pathogenic fungi such as Citrus mould, citrus green mold bacterium, peronophythora litchi bacterium, lichee anthrax bacterias, and active to demonstrating significant anti-corrosive fresh-keeping in the fruit such as lichee, longan, citrus, banana.
For achieving the above object, embodiment of the present invention are: a kind of novel polypeptide antisepsis antistaling agent, are made up of the method comprising the following steps:
(1) be seeded in plain chocolate by the Tibet Kefir granule of late log phase, 30 ~ 40 DEG C of lower seal static fermentations are cultivated 24 ~ 36 hours, produce to part curdled milk;
(2) add and the isopyknic plain chocolate of step (1) gained acidified milk, after rocking, draw liquid fermented milk in an aseptic environment, be coated with respectively with aseptic spreader on MC solid medium and MRS solid state rheology flat board, then seal cultivation;
(3) after cultivating, the bacterium colony in culture dish and culture medium are added in plain chocolate, meanwhile, add Carbon and nitrogen sources, fully stir and each material is mixed, be placed on 30 ~ 35 DEG C of fermentations 65 ~ 75 hours;
(4) after having fermented, the centrifugal 10 ~ 20min of acidified milk, removing precipitation, obtains the supernatant containing novel polypeptide anticorrisive agent;
(5) pH of the supernatant regulated containing natural novel polypeptide anticorrisive agent is 2 ~ 5, keeps 60 ~ 120min in 8 ~ 10 DEG C, and then centrifugal, obtains the precipitation containing novel polypeptide anticorrisive agent;
(6) precipitation is dried to Powdered rear freezen protective, obtains natural novel polypeptide anticorrisive agent;
In step (1), the inoculum concentration of described Tibet Kefir granule in milk is 8 ~ 15%;
As optimization, in step (2), the volume of described absorption liquid fermented milk is 3mL, and the temperature that sealing is cultivated is 28 ~ 35 DEG C.
As optimization, in step (2), add MC solid medium in milk and the dull and stereotyped ratio of MRS solid state rheology is 1:3 ~ 6, the mass ratio of culture medium and milk is 8 ~ 12%.Further optimization, in step (3), add MC solid medium in milk and the dull and stereotyped ratio of MRS solid state rheology is 1:3, the mass ratio of culture medium and milk is 12%,
As optimization, in step (3), the complete mark of described cultivation is that the circular slightly flat single bacterium colony of micro-yellow appears in culture dish surface, and periphery of bacterial colonies produces clear calcium circle.Wherein carbon source is sucrose, glucose and lactose, and quality proportioning is 1 ~ 2:3 ~ 7:0 ~ 3, carbon source and milk mass ratio be 5 ~ 10%, described nitrogenous source is peptone and dusty yeast, and quality proportioning is 3 ~ 5:7 ~ 5, nitrogenous source and milk mass ratio be 4 ~ 8%.Further optimization, the mass volume ratio of described carbon source and milk is 3.84:100, and the quality proportioning of sucrose, glucose, lactose is 2:7:1, and the mass volume ratio of described nitrogenous source and milk is 2.47:100, and the quality proportioning of peptone, dusty yeast is 3:7.The fermentation temperature of further optimization 35 DEG C, fermentation time 75 hours.
As optimization, in step (4), described centrifugation rotating speed is 3000 ~ 5000r/min; The rotating speed of the centrifugation of further optimization is 4500r/min.
As optimization, in step (5), described filtration utilizes 200 ~ 500 object filter clothes to filter; Described pH is 2.2, keeps 120min in 8 ~ 10 DEG C;
As optimization, in step (6), described drying is oven drying or vacuum freeze drying.Further optimization, adopt oven drying, temperature is 50 ± 3 DEG C, and freezen protective temperature is-18 ± 2 DEG C.
The present invention also provides a kind of using method in fruit freshness preserving of novel polypeptide anticorrisive agent, comprise: fruit is soaked in the novel polypeptide anticorrisive agent after being diluted with water, the mass ratio of natural novel polypeptide anticorrisive agent and water is 0.05 ~ 0.3:100, pull out after first soaking 5 ~ 15min, then drain, normal temperature storage or refrigeration.
The present invention confirms that the novel polypeptide aseptic active obtained detects through the experiment of Oxford cup and analyzes, and finds that it has significant inhibition to fruit spoilage organisms such as staphylococcus aureus, salmonella, Escherichia coli with to fruit postharvest pathogenic fungi such as Citrus mould, citrus green mold bacterium, peronophythora litchi bacterium, lichee anthrax bacterias.Meanwhile, it is to light, thermally-stabilised, and long shelf-life, for its extensive use in the fresh-keeping field of fruit antisepsis is had laid a good foundation.
The invention has the advantages that: the novel polypeptide anticorrisive agent of (1) gained of the present invention is that separation and purification obtains from the acidified milk by Tibet Kefir grains, safety and sanitation, Nantural non-toxic, effectively reduce chemical agent to the harm of the mankind; (2) production technology of the novel polypeptide anticorrisive agent of gained of the present invention is simple, and easy and simple to handle, output is high, and fermentation medium main component is plain chocolate, and cost is low, is suitable for industrialization and produces, have significant economic benefit; (3) the novel polypeptide anticorrisive agent of gained of the present invention has significant inhibition, has a broad antifungal spectrum to fruit spoilage organisms such as staphylococcus aureus, salmonella, Escherichia coli with to fruit postharvest pathogenic fungi such as Citrus mould, citrus green mold bacterium, peronophythora litchi bacterium, lichee anthrax bacterias.Meanwhile, it is to light, thermally-stabilised, and long shelf-life, makes it have very large potential using value in the fresh-keeping field of fruit antisepsis.(4) the novel polypeptide anticorrisive agent of gained of the present invention can preserve nutritional labeling and the mouthfeel of fruit to greatest extent, and play good effect of color protection, extend its shelf life, simultaneously also for the technical barrier that fruit postharvest diseases generation prevented and treated by main dependence chemical bactericide at present provides reference.
Detailed description of the invention
Provide embodiments of the invention below, specifically set forth the present invention.
embodiment 1:
The preparation method of novel polypeptide antisepsis antistaling agent:
Tibet Kefir granule is seeded to plain chocolate after 30 DEG C of lower seal static fermentations cultivate 24 hours, produces to part curdled milk.Then add the isopyknic plain chocolate of acidified milk, after rocking, draw 3mL liquid fermented milk in an aseptic environment, be coated with respectively with aseptic spreader on MC solid medium and MRS solid state rheology flat board and cultivate in 30 DEG C of sealings.Treat that the circular slightly flat single bacterium colony of micro-yellow appears in culture dish surface, when periphery of bacterial colonies produces clear calcium circle, the bacterium colony in culture dish and culture medium to be added in plain chocolate and to add carbon source and nitrogenous source, in 35 DEG C of fermentation times 75 hours.Wherein, MC solid medium is dull and stereotyped is 1:3 with the number ratio of MRS solid state rheology flat board, the mass ratio of culture medium and milk is 12%, the mass volume ratio of carbon source and milk is 3.84:100, the ratio of sucrose, glucose, lactose is 2:7:1, the mass volume ratio of nitrogenous source and milk is 2.47:100, and the ratio of peptone, dusty yeast is 3:7.After having fermented, the centrifugal 10 ~ 20min of acidified milk 4500r/min, removing precipitation, obtains the supernatant containing novel polypeptide anticorrisive agent.Then the pH of the supernatant containing natural novel polypeptide anticorrisive agent is regulated to be 2.2 and to keep 120min in 8 ~ 10 DEG C, more centrifugal, obtain the precipitation containing novel polypeptide anticorrisive agent.Finally, will precipitate and 50 DEG C of oven dryings, just obtain the powder of natural novel polypeptide anticorrisive agent.
Tibet Kefir granule is seeded to plain chocolate after 30 DEG C of lower seal static fermentations cultivate 24 hours, produces to part curdled milk.Then add the isopyknic plain chocolate of acidified milk, after rocking, draw 3mL liquid fermented milk in an aseptic environment, be coated with respectively with aseptic spreader on MC solid medium and MRS solid state rheology flat board and cultivate in 30 DEG C of sealings.Treat that the circular slightly flat single bacterium colony of micro-yellow appears in culture dish surface, when periphery of bacterial colonies produces clear calcium circle, the bacterium colony in culture dish and culture medium to be added in plain chocolate and to add carbon source and nitrogenous source, in 32 DEG C of fermentation times 80 hours.Wherein, MC solid medium is dull and stereotyped is 1:3 with the number ratio of MRS solid state rheology flat board, the mass ratio of culture medium and milk is 13%, the mass volume ratio of carbon source and milk is 3.84:100, the ratio of sucrose, glucose, lactose is 2:7:1, the mass volume ratio of nitrogenous source and milk is 2.47:100, and the ratio of peptone, dusty yeast is 3:7.After having fermented, the centrifugal 10 ~ 20min of acidified milk 4500r/min, removing precipitation, obtains the supernatant containing novel polypeptide anticorrisive agent.Then the pH of the supernatant containing natural novel polypeptide anticorrisive agent is regulated to be 2.2 and to keep 120min in 8 ~ 10 DEG C, more centrifugal, obtain the precipitation containing novel polypeptide anticorrisive agent.Finally, will precipitate and 50 DEG C of oven dryings, just obtain the powder of natural novel polypeptide anticorrisive agent.
Tibet Kefir granule is seeded to plain chocolate after 30 DEG C of lower seal static fermentations cultivate 24 hours, produces to part curdled milk.Then add the isopyknic plain chocolate of acidified milk, after rocking, draw 3mL liquid fermented milk in an aseptic environment, be coated with respectively with aseptic spreader on MC solid medium and MRS solid state rheology flat board and cultivate in 30 DEG C of sealings.Treat that the circular slightly flat single bacterium colony of micro-yellow appears in culture dish surface, when periphery of bacterial colonies produces clear calcium circle, the bacterium colony in culture dish and culture medium to be added in plain chocolate and to add carbon source and nitrogenous source, in 36 DEG C of fermentation times 73 hours.Wherein, MC solid medium is dull and stereotyped is 1:3 with the number ratio of MRS solid state rheology flat board, the mass ratio of culture medium and milk is 15%, the mass volume ratio of carbon source and milk is 3.84:100, the ratio of sucrose, glucose, lactose is 2:7:1, the mass volume ratio of nitrogenous source and milk is 2.47:100, and the ratio of peptone, dusty yeast is 3:7.After having fermented, the centrifugal 10 ~ 20min of acidified milk 4500r/min, removing precipitation, obtains the supernatant containing novel polypeptide anticorrisive agent.Then the pH of the supernatant containing natural novel polypeptide anticorrisive agent is regulated to be 2.2 and to keep 120min in 8 ~ 10 DEG C, more centrifugal, obtain the precipitation containing novel polypeptide anticorrisive agent.Finally, will precipitate and 50 DEG C of oven dryings, just obtain the powder of natural novel polypeptide anticorrisive agent.
embodiment 2:
Novel polypeptide antisepsis antistaling agent provided by the invention is to fruit spoilage organisms and fruit postharvest pathogenic fungi inhibition, and adopt Oxford cup agar diffusion method to analyze novel polypeptide antisepsis antistaling agent, concrete grammar is:
2.1 indicator bacteria activation
The single bacterium colony (fruit spoilage organisms) of the dull and stereotyped indicator bacteria of the test tube that picking 4 DEG C is preserved in superclean bench in 7mLLB broth bouillon, is positioned over constant-temperature table, 150r/min incubated overnight at 37 DEG C.It is 1 × 10 that the indicator bacteria liquid of activated overnight is diluted to bacteria concentration
8cFU/mL, for subsequent use.
In superclean bench, the single bacterium colony (fruit postharvest pathogenic fungi) of the dull and stereotyped indicator bacteria of the test tube of picking 4 DEG C preservation is in ruling at PDA solid medium, constant incubator incubated overnight at being positioned over 30 DEG C.By the indicator bacteria liquid of activated overnight, transfer a single bacterium colony and shake up in the sterilized water of 0.2mL, bacterium colony is uniformly dispersed, for subsequent use.
2.2 detect dull and stereotyped preparation
The LB agar of sterilization treatment is cooled to 50 ~ 60 DEG C, under aseptic condition, in the culture dish (9cm) of horizontal positioned, shifts 15mL culture medium with 5mL pipettor.After agar plate solidifies, draw 0.2mL indicator bacteria liquid (fruit spoilage organisms) to dull and stereotyped, even with aseptic spreader coating, place sterilized Oxford cup (internal diameter 6 ± 0.1mm, external diameter 7.8 ± 0.1mm, high 10 ± 0.1mm) subsequently on culture dish flat board.For preventing Oxford cup from placing built on the sand, can suitably press.
The PDA agar of sterilization treatment is cooled to 50 ~ 60 DEG C, under aseptic condition, in the culture dish (9cm) of horizontal positioned, shifts 15mL culture medium with 5mL pipettor.After agar plate solidifies, draw 0.2mL indicator bacteria liquid (fruit postharvest pathogenic fungi) to dull and stereotyped, even with aseptic spreader coating, place sterilized Oxford cup (internal diameter 6 ± 0.1mm subsequently, external diameter 7.8 ± 0.1mm, high 10 ± 0.1mm) on culture dish flat board.For preventing Oxford cup from placing built on the sand, can suitably press.
2.3 Antibacterial Activity
0.2mL novel polypeptide anti-corrosive fresh-keeping agent solution (100mg/mL) is accurately drawn to Oxford cup with liquid-transfering gun, containing flat board constant temperature quiescent culture 412h at 37 DEG C of fruit spoilage organisms, and containing flat board constant temperature quiescent culture 48 ~ 72h at 30 DEG C of fruit postharvest pathogenic fungi, antibacterial circle diameter surveyed by slide measure.Taken pictures from culture dish back scan by scanner.Novel polypeptide antisepsis antistaling agent to fruit spoilage organisms and fruit postharvest pathogenic fungi inhibition in table 1.
The antibacterial effect (X ± SD, n=3) of table 1 novel polypeptide antisepsis antistaling agent
| Indicator bacteria | Antibacterial activity |
| Escherichia coli | +++ |
| Staphylococcus aureus | +++ |
| Salmonella | +++ |
| Bacillus thuringiensis | +++ |
| Shigella dysenteriae | +++ |
| Citrus becomes sour germ | +++ |
| Citrus pathogens penicillium | + |
| Penicillium digitatum | ++ |
| Loquat anthrax bacteria | ++ |
| Loquat Leaf Spot bacterium | +++ |
| Loquat alternaria | ++ |
| Banana crown rot bacterium | ++ |
| Glorosprium musarum Cookeet Mass | ++ |
| Lichee anthrax bacteria | +++ |
| Peronophythora Litchii germ | + |
Note :+: inhibition zone 5 ~ 10mm; ++: inhibition zone 10 ~ 20mm; +++: inhibition zone >20mm
Can know from table 1, the antimicrobial spectrum of novel polypeptide antisepsis antistaling agent is relatively extensive, and this has significant inhibition to fruit spoilage organisms such as staphylococcus aureus, salmonella, Escherichia coli with to fruit postharvest pathogenic fungi such as Citrus mould, citrus green mold bacterium, peronophythora litchi bacterium, lichee anthrax bacterias.
embodiment 3:
The stability of novel polypeptide antisepsis antistaling agent provided by the invention measures:
The photostability of 3.1 novel polypeptide antisepsis antistaling agents measures
After different light process 60min, the antibacterial activity of novel polypeptide antisepsis antistaling agent is as shown in table 2, and the fungistatic effect of different light medium on novel polypeptide antisepsis antistaling agent does not almost affect, and illustrates that novel polypeptide antisepsis antistaling agent has good photostability.
The fungistatic effect of table 2 novel polypeptide antisepsis antistaling agent after different light process 60min
| Contrast | Lucifuge | Incandescent light | Ultraviolet light | Infrared light | |
| Staphylococcus aureus | +++ | +++ | +++ | +++ | +++ |
| Salmonella | +++ | +++ | +++ | +++ | +++ |
| Citrus pathogens penicillium | + | + | + | + | + |
| Loquat alternaria | ++ | ++ | ++ | ++ | ++ |
| Banana crown rot bacterium | ++ | ++ | ++ | ++ | ++ |
| Peronophythora Litchii germ | + | + | + | + | + |
Note :+: inhibition zone 5 ~ 10mm; ++: inhibition zone 10 ~ 20mm; +++: inhibition zone >20mm
The thermal stability determination of 3.2 novel polypeptide antisepsis antistaling agents
After heat treatment, the change of novel polypeptide antisepsis antistaling agent antibacterial effect is as shown in table 3, novel polypeptide antisepsis antistaling agent its fungistatic effect after-20 DEG C, 4 DEG C, 40 DEG C, 70 DEG C, 90 DEG C, 121 DEG C process 15min is almost unchanged, after 121 DEG C of high-temperature process 15min, novel polypeptide antisepsis antistaling agent does not have yet indicator bacteria bacteriostasis and obviously weakens, this explanation, novel polypeptide antisepsis antistaling agent energy higher temperature resistant is more stable to heat.
The fungistatic effect of table 3 novel polypeptide antisepsis antistaling agent after treatment of different temperature 15min
| Normal temperature | -20℃ | 4℃ | 40℃ | 70℃ | 90℃ | 121℃ | |
| Staphylococcus aureus | +++ | +++ | +++ | +++ | +++ | +++ | ++ |
| Salmonella | +++ | +++ | +++ | +++ | +++ | +++ | +++ |
| Citrus pathogens penicillium | + | + | + | + | + | + | + |
| Loquat alternaria | ++ | ++ | ++ | ++ | ++ | ++ | + |
| Banana crown rot bacterium | ++ | ++ | ++ | ++ | ++ | ++ | ++ |
| Peronophythora Litchii germ | + | + | + | + | + | + | + |
Note :+: inhibition zone 5 ~ 10mm; ++: inhibition zone 10 ~ 20mm; +++: inhibition zone >20mm
The thermal stability determination of 3.3 novel polypeptide antisepsis antistaling agents
As can be seen from Table 4, after novel polypeptide antisepsis antistaling agent preserves 360d under normal temperature condition, it does not weaken the bacteriostatic activity of indicator bacteria, do not have significant difference (
p>0.05), show that the stability of novel polypeptide antisepsis antistaling agent is better, can preserve for a long time.
The impact of number of days on the fungistatic effect of novel polypeptide antisepsis antistaling agent preserved by table 4
| Preserve 0d | Preserve 60d | Preserve 120d | Preserve 240d | Preserve 360d | |
| Staphylococcus aureus | +++ | +++ | +++ | +++ | +++ |
| Salmonella | +++ | +++ | +++ | +++ | +++ |
| Citrus pathogens penicillium | + | + | + | + | + |
| Loquat alternaria | ++ | ++ | ++ | ++ | ++ |
| Banana crown rot bacterium | ++ | ++ | ++ | ++ | ++ |
| Peronophythora Litchii germ | + | + | + | + | + |
Note :+: inhibition zone 5 ~ 10mm; ++: inhibition zone 10 ~ 20mm; +++: inhibition zone >20mm
In sum, novel polypeptide antisepsis antistaling agent is to light, thermally-stabilised, and long shelf-life, for its extensive use in the fresh-keeping field of fruit antisepsis is had laid a good foundation.
embodiment 3:
Novel polypeptide antisepsis antistaling agent provided by the invention is tested the keeping-freshness storage of fruit:
3.1 buy " osmanthus taste " fresh lichees, reject wormed fruit, select size homogeneous, without stain, have no mechanical damage stay base of a fruit lichee, with the clean rear natural air drying of clean water, stochastic averagina is divided into 4 groups.Use 0.05%FX-6 extractive from fermentative, 0.1%FX-6 extractive from fermentative, 0.05% P applied levels and distilled water immersion 2min respectively, ventilation Quick-air-drying is placed on after picking up lichee, put down gently in bubble chamber, ensure that lichee is not extruded, good seal is placed in the thermostatic chamber preservation of normal temperature (25 DEG C ± 3 DEG C), regularly detects the physicochemical property etc. of lichee.
3.1.1 the mensuration of storage effect
From the lichee after process, often group chooses much the same 100 lichee of size, loads sealed polyethylene plastic bag, for the detection of mass loss rate, browning index and healthy fruit.
Wherein:
Once quality × 100% is claimed before mass loss rate=(once claiming quality after once claiming quality-fruit before fruit)/fruit.
Brown stain grade scale: 1 grade of fruit is less than 1/4 of pericarp area without brown stain or browned area; 2 grades of fruit browned area account for pericarp area 1/4 ~ 1/2; 3 grades of fruit browned area account for pericarp area 1/2 ~ 3/4; 4 grades of fruit browned area are greater than 3/4 of pericarp area; 5 grades of fruits are complete brown stain.
Browning index=∑ (brown stain progression × brown stain fruit number at different levels)/test fruit sum.
By pericarp browning progression be 1 and 2 fruit be decided to be fruit.
Healthy fruit=(1 grade of fruit number+2 grades of fruit number)/investigation fruit total quantity × 100%.
As shown in Table 5, our known 0.05%, 0.1% novel polypeptide antisepsis antistaling agent can reduce the moisture loss of fruit tissue at duration of storage effectively; 0.05%, 0.1% novel polypeptide antisepsis antistaling agent can suppress litchi fruits pericarp browning, maintain the good appearance luster of litchi fruits; 0.05%, 0.1% novel polypeptide antisepsis antistaling agent has good fresh-keeping effect, and after lichee normal temperature storage 5d, healthy fruit also has more than 88%.
The change of table 5 litchi fruits mass loss rate, browning index and healthy fruit
3.1.2 the mensuration of lichee quality
10 lichee are randomly drawed, by the firmness change of instrumental test duration of storage litchi pulp from each process; From each process, randomly draw 12 fruits, broken through bruisher, low-temperature centrifugation, gets supernatant and carries out soluble solid, titrable acidity, content of reducing sugar, ascorbic mensuration.Wherein, soluble solid content digital display refractometer measures, and each parallel determination 10 times, averages.Titrable acidity adopts determination of acid-basetitration; 3,5-dinitrosalicylic acid system measures content of reducing sugar; HPLC method measures the content of ascorbic acid; Above Indexs measure result is the mean value of 3 parallel determinations.
By the change of the table 6 litchi fruits index of quality, we can know: 0.05%, 0.1% novel polypeptide antisepsis antistaling agent can suppress the activity of respiration and cell wall degrading enzyme, the decline suppressing the hardness of fruit, delayed fruit Ripening and Softening; 0.05%, 0.1% novel polypeptide antisepsis antistaling agent can suppress soluble solid, content of reducing sugar, titratable acid, ascorbic consumption preferably, thus maintains local flavor and the nutritional quality of lichee to greatest extent, reaches good fresh-keeping effect.
The change of the table 6 litchi fruits index of quality
The fresh-keeping test of 3.2 other fruit
Tune fresh, size evenly, have no mechanical damage, after strawberry wash clean without disease and pest, after being immersed in the novel polypeptide antisepsis antistaling agent aqueous solution of 0.04% 5 minutes, taking out, draining, to be placed under normal temperature (25 ± 3 DEG C) fresh-keeping 4 days, healthy fruit more than 94%.
Select size Nanfeng orange that is even, that damage without disease and pest, maturity consistent (medium well), surface inorganic tool, first use clean water, after natural air drying, to be immersed in the novel polypeptide antisepsis antistaling agent aqueous solution of 0.07% 10 minutes, after taking out, draining, load sealed polyethylene plastic bag, be placed in the Cold storage in the refrigerator of 4 ± 2 DEG C, after 100 days, its healthy fruit is more than 93.2%.
Buy fresh longan, reject wormed fruit, select size homogeneous, without stain, have no mechanical damage stay Tyrone eye, natural air drying after clean by clean water, after being immersed in the novel polypeptide antisepsis antistaling agent aqueous solution of 0.05% 2 minutes, taking out, draining, load sealed polyethylene plastic bag, to be placed under normal temperature (25 ± 3 DEG C) fresh-keeping 8 days, healthy fruit more than 95%.
In sum, the novel polypeptide antisepsis antistaling agent of gained of the present invention can solve the fresh-keeping problem of the fruit such as lichee, strawberry, Nanfeng orange, longan well, reaches good corrosion-resistanting fresh-keeping effect, has great application prospect.
Claims (15)
1. a preparation method for novel polypeptide antisepsis antistaling agent, is characterized in that, comprises the following steps:
(1) be seeded in plain chocolate by the Tibet Kefir granule of late log phase, 30 ~ 40 DEG C of lower seal static fermentations are cultivated 24 ~ 36 hours, produce to part curdled milk;
(2) add and the isopyknic plain chocolate of step (1) gained acidified milk, after rocking, draw liquid fermented milk in an aseptic environment, be coated with respectively with aseptic spreader on MC solid medium and MRS solid state rheology flat board, then seal cultivation;
(3) after cultivating, the bacterium colony in culture dish and culture medium are added in plain chocolate, meanwhile, add Carbon and nitrogen sources, fully stir and each material is mixed, be placed on 30 ~ 35 DEG C of fermentations 65 ~ 75 hours;
(4) after having fermented, the centrifugal 10 ~ 20min of acidified milk, removing precipitation, obtains the supernatant containing novel polypeptide anticorrisive agent;
(5) pH of the supernatant regulated containing natural novel polypeptide anticorrisive agent is 2 ~ 5, keeps 60 ~ 120min in 8 ~ 10 DEG C, and then centrifugal, obtains the precipitation containing novel polypeptide anticorrisive agent;
(6) precipitation is dried to Powdered rear freezen protective, obtains natural novel polypeptide anticorrisive agent.
2. the preparation method that obtains of novel polypeptide antisepsis antistaling agent according to claim 1, it is characterized in that, in step (1), the inoculum concentration of described Tibet Kefir granule in milk is 8 ~ 15%.
3. the preparation method of the novel polypeptide antisepsis antistaling agent according to claim, is characterized in that, in step (2), the volume of described absorption liquid fermented milk is 3mL, and the temperature that sealing is cultivated is 28 ~ 35 DEG C.
4. the preparation method that the novel polypeptide antisepsis antistaling agent stated according to claim 1 obtains, is characterized in that, in step (2), add MC solid medium in milk and the dull and stereotyped ratio of MRS solid state rheology is 1:3 ~ 6, the mass ratio of culture medium and milk is 8 ~ 12%.
5. the preparation method of the novel polypeptide antisepsis antistaling agent according to claim 1 or 3, is characterized in that, in step (3), add MC solid medium in milk and the dull and stereotyped ratio of MRS solid state rheology is 1:3, the mass ratio of culture medium and milk is 12%.
6. the preparation method of novel polypeptide antisepsis antistaling agent according to claim 1, is characterized in that, in step (3), the complete mark of described cultivation is that the circular slightly flat single bacterium colony of micro-yellow appears in culture dish surface, and periphery of bacterial colonies produces clear calcium circle.
7. the preparation method of novel polypeptide antisepsis antistaling agent according to claim 1, it is characterized in that, in step (3), carbon source is sucrose, glucose and lactose, quality proportioning is 1 ~ 2:3 ~ 7:0 ~ 3, carbon source and milk mass ratio be 5 ~ 10%, described nitrogenous source is peptone and dusty yeast, quality proportioning is 3 ~ 5:7 ~ 5, nitrogenous source and milk mass ratio be 4 ~ 8%.
8. the preparation method of the novel polypeptide antisepsis antistaling agent according to claim 1 or 6, it is characterized in that, described in step (3), the mass volume ratio of carbon source and milk is 3.84:100, the quality proportioning of sucrose, glucose, lactose is 2:7:1, the mass volume ratio of described nitrogenous source and milk is 2.47:100, and the quality proportioning of peptone, dusty yeast is 3:7.
9. the preparation method that the novel polypeptide antisepsis antistaling agent stated according to claim 1 obtains, is characterized in that, fermentation temperature 35 DEG C, fermentation time 75 hours.
10. the preparation method that the novel polypeptide antisepsis antistaling agent stated according to claim 1 obtains, is characterized in that, in step (4), described centrifugation rotating speed is 3000 ~ 5000r/min.
The preparation method that the 11. novel polypeptide antisepsis antistaling agents stated according to claim 1 obtain, it is characterized in that, in step (5), described filtration utilizes 200 ~ 500 object filter clothes to filter.
The preparation method that the 12. novel polypeptide antisepsis antistaling agents stated according to claim 1 obtain, it is characterized in that, in step (5), described pH is 2.2, keeps 120min1 in 8 ~ 10 DEG C.
The preparation method that the 13. novel polypeptide antisepsis antistaling agents stated according to claim 1 obtain, it is characterized in that, adopt oven drying, temperature is 50 ± 3 DEG C, and freezen protective temperature is-18 ± 2 DEG C.
Polypeptide antisepsis antistaling agent prepared by 14. 1 kinds of methods as described in claim 1-13 any one.
The using method of 15. 1 kinds of polypeptide antisepsis antistaling agents as claimed in claim 13, it is characterized in that, comprise: fruit is soaked in the novel polypeptide anticorrisive agent after being diluted with water, the mass ratio of natural novel polypeptide anticorrisive agent and water is 0.05 ~ 0.3:100, pull out after first soaking 5 ~ 15min, then drain, normal temperature storage or refrigeration.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109827920A (en) * | 2018-12-29 | 2019-05-31 | 南京同凯兆业生物技术有限责任公司 | The detection method of reduced sugar in a kind of Penicillium citrinum fermentation liquid |
| CN110574783A (en) * | 2019-09-20 | 2019-12-17 | 华南农业大学 | Fruit cucumber preservation method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104087524A (en) * | 2014-04-02 | 2014-10-08 | 曹庸 | Lactobacillus paracasei subsp. tolerans strain, screening method and use |
| CN104855492A (en) * | 2015-04-02 | 2015-08-26 | 曹庸 | A natural preservative for meat products and a preparation and use method thereof |
-
2015
- 2015-12-16 CN CN201510937458.4A patent/CN105532855A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104087524A (en) * | 2014-04-02 | 2014-10-08 | 曹庸 | Lactobacillus paracasei subsp. tolerans strain, screening method and use |
| CN104855492A (en) * | 2015-04-02 | 2015-08-26 | 曹庸 | A natural preservative for meat products and a preparation and use method thereof |
Non-Patent Citations (3)
| Title |
|---|
| 卢曼等: "西藏开菲尔粒中酵母菌的分离与初步鉴定", 《上海海洋大学学报》 * |
| 苗建银等: "西藏开菲尔发酵提取物对牛奶和馒头防腐作用的研究", 《现代食品科技》 * |
| 黄曼曼等: "西藏开菲尔中抑菌活性物质的提取及抑菌效果研究", 《食品工业科技》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109827920A (en) * | 2018-12-29 | 2019-05-31 | 南京同凯兆业生物技术有限责任公司 | The detection method of reduced sugar in a kind of Penicillium citrinum fermentation liquid |
| CN110574783A (en) * | 2019-09-20 | 2019-12-17 | 华南农业大学 | Fruit cucumber preservation method |
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