CN102630746B - Yeast composite biological preservative and preparation method and application thereof - Google Patents
Yeast composite biological preservative and preparation method and application thereof Download PDFInfo
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- CN102630746B CN102630746B CN 201210143690 CN201210143690A CN102630746B CN 102630746 B CN102630746 B CN 102630746B CN 201210143690 CN201210143690 CN 201210143690 CN 201210143690 A CN201210143690 A CN 201210143690A CN 102630746 B CN102630746 B CN 102630746B
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Abstract
The invention discloses a yeast composite biological preservative and a preparation method and application of the preservative. Every 100mL of yeast composite biological preservative contains 1*10<10> CFU (colony forming unit) of Pichia membranaefaciens, 0-2g of chitosan oligosaccharide, 0.5-1g of chitosan, 1-2g of calcium chloride, 0.5mL of acetic acid and the balance of water, and the pH value is 5.8 to 6. The yeast composite biological preservative is used for dipping treatment of citrus fruits, and can be used to reduce the disease rate of harvested citrus fruits significantly, delay the fruit senescence, and prolong the storage period and shelf life of the fruits. The problems of possible chemical residue and environment pollution caused by existing chemical microbicide for fresh-keeping of citrus fruits are avoided. The preparation method is simple, low in cost and easy to popularize and apply.
Description
Technical field
The invention belongs to the fruit freshness preserving technical field, relate to a kind of fruit antistaling agent and its preparation method and application.
Background technology
Oranges and tangerines (citrus fruits) are the big fruit of the first in the world, the second largest fruit of China, occupy very consequence in China's agricultural economy, have become leading industry in many areas.But, to compare with the oranges and tangerines plant husbandry of fast development, the technical research in postpartum of oranges and tangerines fresh fruit and industrialized development are relatively slow, and be annual preserving and all causing a large amount of fruit rots rotten between the delivery period, causes significant wastage and the serious economy loss of resource.
After citrusfruit is gathered, more than 20 kind of postharvest disease can take place in transporting procedures, mainly be divided into physiological disturbance and infectious disease two big classes.Physiological disturbance mainly contains oil mark disease, brown spot, oedema and low water disease etc.Mainly contain penicilliosis, green mould, sour rot, anthracnose, blossom-end rot and brown rot etc. by fungus-caused infectious disease.Wherein to infect rotting of causing be to adopt the huge main reason of back loss to fungal disease.At present, the postharvest disease means of prevention of citrusfruit mainly is that deepfreeze and chemical bactericide are handled.Deepfreeze can reduce the respiratory metabolism of fruit and the rotting rate of fruit, but equipment needed thereby cost and energy consumption are higher.Chemical bactericide can be killed pathogen, but use the resistance increase that has made pathogen for years and cause prevention effect to reduce, the growing interest of and food-safety problem healthy to self along with people simultaneously, problems such as the medicament residue that the use chemical bactericide may relate to and environmental pollution, also make the use that is under an embargo gradually of some effective chemical bactericide, these have all weakened the ability that people control the oranges and tangerines postharvest disease greatly.Therefore need seek the new antistaling agent of citrusfruit safely and efficiently.
In recent years many studies show that can utilize fruit and vegetable surfaces to have the beneficial microbe (as bacterium, saccharomycete, small-sized filamentous fungi etc.) of antagonism to suppress growth of pathogenic bacteria to pathogen, prevents from adopting the generation of back fungal disease.Wherein, antagonism yeast good owing to antagonistic effect, do not produce the main bacteria seed that advantage such as antibiotic becomes biologically controlling postharvest disease research.But less about the research of antagonism yeast control citrusfruit postharvest disease at present, more lack antagonism yeast and other composite preservative research of bactericide class material non-chemically.
Summary of the invention
In view of this, one of purpose of the present invention is to provide a kind of safe, yeast complex biological preservative efficiently, two of purpose is to provide the preparation method of described yeast complex biological preservative, three of purpose is to provide the application of described yeast complex biological preservative in citrusfruit is fresh-keeping, with control disease, the delayed fruit aging of citrusfruit storage period, avoid simultaneously existing for the fresh-keeping chemical bactericide of citrusfruit may problem, the problem includes: problems such as medicament residue and environmental pollutions.
For achieving the above object, the invention provides following technical scheme:
1. the yeast complex biological preservative contains 1 * 10 among every 100mL
10The CFU Pichia membranaefaciens (
Pichia membranaefaciens), 0 ~ 2g chitosan oligosaccharide, 0.5 ~ 1g shitosan, 1 ~ 2g calcium chloride and 0.5mL acetic acid, surplus is water, regulating the pH value is 5.8-6.
Preferably, contain 1 * 10 among the every 100mL of described yeast complex biological preservative
10CFU Pichia membranaefaciens, 0.5g shitosan, 1g calcium chloride and 0.5mL acetic acid, surplus are water, and regulating the pH value is 5.8-6.
Preferably, contain 1 * 10 among the every 100mL of described yeast complex biological preservative
10CFU Pichia membranaefaciens, 2g chitosan oligosaccharide, 0.5g shitosan, 1g calcium chloride and 0.5mL acetic acid, surplus are water, and regulating the pH value is 5.8-6.
2. the preparation method of described yeast complex biological preservative may further comprise the steps: add acetic acid and water in Pichia membranaefaciens suspension, make every 100mL and contain 1 * 10
10The mixed liquor of CFU yeast and 0.5mL acetic acid adds chitosan oligosaccharide, shitosan and calcium chloride again in this mixed liquor, make among every 100mL and contain 1 * 10
10The mixed liquor of CFU Pichia membranaefaciens, 0 ~ 2g chitosan oligosaccharide, 0.5 ~ 1g shitosan, 1 ~ 2g calcium chloride and 0.5mL acetic acid, regulating the pH value is 5.8-6, namely gets the yeast complex biological preservative.
Further, the preparation method of described Pichia membranaefaciens suspension is: Pichia membranaefaciens is inoculated in the NYDB nutrient solution, and in 28 ± 1 ℃ of shaken cultivation of temperature, the centrifugal supernatant of abandoning, it is resuspended that yeast cells adds water, namely gets Pichia membranaefaciens suspension.
3. the application of described yeast complex biological preservative in citrusfruit is fresh-keeping.
4. utilize described yeast complex biological preservative to carry out the fresh-keeping method of citrusfruit: citrusfruit was soaked 2 minutes in described yeast complex biological preservative.
Beneficial effect of the present invention is: the invention provides a kind of safe, yeast complex biological preservative and preparation method thereof and application in citrusfruit is fresh-keeping efficiently, with this yeast complex biological preservative citrusfruit is carried out immersion treatment, can significantly reduce the generation of citrusfruit postharvest disease, the delayed fruit aging, prolong storage period and the shelf life of fruit, avoid existing for the fresh-keeping chemical bactericide of citrusfruit may problem, the problem includes: problems such as medicament residue and environmental pollutions, and its preparation method is simple, cost is low, is easy to apply.
Description of drawings
In order to make the purpose, technical solutions and advantages of the present invention clearer, the present invention is described in further detail below in conjunction with accompanying drawing, wherein:
Fig. 1 is yeast and shitosan, CaCl
2Combined Processing is to the influence of the citrusfruit green mold incidence of disease.
Fig. 2 is yeast and shitosan, CaCl
2Combined Processing is to the influence of citrusfruit green mould spot diameter.
Fig. 3 is yeast and chitosan oligosaccharide, shitosan, CaCl
2Combined Processing is to the influence of citrusfruit natural occurrence rate.
Fig. 4 is yeast and chitosan oligosaccharide, shitosan, CaCl
2Combined Processing is to the influence of the common storage disease natural occurrence of citrusfruit rate, and wherein scheming A is each disease incidence of preceding 20 days of storage period, and figure B is each disease incidence of whole duration of storage (60 days).
Fig. 5 is yeast and chitosan oligosaccharide, shitosan, CaCl
2Combined Processing is to the influence of citrusfruit weight-loss ratio.
Fig. 6 is yeast and chitosan oligosaccharide, shitosan, CaCl
2Combined Processing is to the influence of citrusfruit soluble solid content.
Fig. 7 is yeast and chitosan oligosaccharide, shitosan, CaCl
2Combined Processing is to the influence of citrusfruit titratable acid content.
Fig. 8 is yeast and chitosan oligosaccharide, shitosan, CaCl
2Combined Processing is to the influence of citrusfruit ascorbic acid content.
The specific embodiment
Below with reference to accompanying drawings the preferred embodiments of the present invention are described in detail.
One, experiment material
The kind of oranges and tangerines described in the preferred embodiment is northern a word used in place name 447
#The brocade orange (
Citrus SinensicOsbeck. cv. Jincheng 447
#), pick up from gold orchard, Beibei District, Chongqing City.Select fresh nothing machinery and hinder, big or small homogeneous, the fruit of maturity unanimity is as experiment material.Fruit is adopted the back, and storage is standby down at 4 ℃.
The green mold bacterium (
Penicillium digitatumSacc.) preserve bacterial classification for this laboratory.Pichia membranaefaciens (
Pichia membranaefaciens) be the yeast for brewing litchi wine, available from Chinese common micro-organisms culture presevation administrative center; Connect this yeast of ring in 20 ml NYDB nutrient solution (yeast extract 5g, beef extract 8g, glucose 10g from slant medium during use, water 1000ml) in, 28 ℃ of following shaken cultivation 24 hours, under 6000 r/min centrifugal 10 minutes, abandon supernatant, add sterilized water, make 1 * 10 with hemocytometer
8The CFU/ml cell suspending liquid.
Two, yeast and shitosan, CaCl
2
Combined Processing is to the influence of citrusfruit green mold disease
Prescription A1:0.1% (g/ml) shitosan+1 * 10
8The CFU/ml yeast cream;
Prescription A2:0.5% (g/ml) shitosan+1 * 10
8The CFU/ml yeast cream;
Prescription A3:1.0% (g/ml) shitosan+1 * 10
8The CFU/ml yeast cream;
Prescription B1:0.5% (g/ml) CaCl
2+ 1 * 10
8The CFU/ml yeast cream;
Prescription B2:1.0% (g/ml) CaCl
2+ 1 * 10
8The CFU/ml yeast cream;
Prescription B3:2.0% (g/ml) CaCl
2+ 1 * 10
8The CFU/ml yeast cream;
Prescription C1:0.1% (g/ml) shitosan+0.5% (g/ml) CaCl
2+ 1 * 10
8The CFU/ml yeast cream;
Prescription C2:0.1% (g/ml) shitosan+1.0% (g/ml) CaCl
2+ 1 * 10
8The CFU/ml yeast cream;
Prescription C3:0.1% (g/ml) shitosan+2.0% (g/ml) CaCl
2+ 1 * 10
8The CFU/ml yeast cream;
Prescription D1:0.5% (g/ml) shitosan+0.5% (g/ml) CaCl
2+ 1 * 10
8The CFU/ml yeast cream;
Prescription D2:0.5% (g/ml) shitosan+1.0% (g/ml) CaCl
2+ 1 * 10
8The CFU/ml yeast cream;
Prescription D3:0.5% (g/ml) shitosan+2.0% (g/ml) CaCl
2+ 1 * 10
8The CFU/ml yeast cream;
Prescription E1:1.0% (g/ml) shitosan+0.5% (g/ml) CaCl
2+ 1 * 10
8The CFU/ml yeast cream;
Prescription E2:1.0% (g/ml) shitosan+1.0% (g/ml) CaCl
2+ 1 * 10
8The CFU/ml yeast cream;
Prescription E3:1.0% (g/ml) shitosan+2.0% (g/ml) CaCl
2+ 1 * 10
8The CFU/ml yeast cream;
Preparation method: to 1 * 10
8Adding acetic acid to final concentration in the CFU/ml yeast cream is 0.5% (ml/ml), adds certain amount of chitosan, CaCl again
2To final concentration shown in above-mentioned prescription A1 ~ A3, B1 ~ B3, C1 ~ C3, D1 ~ D3, the E1 ~ E3, regulate the pH value to 5.8-6, namely with the 0.1mol/L sodium hydroxide solution at last.
Get the surface through the citrusfruit of 75% (ml/ml) alcohol wipe sterilization, with the sterilization after prick punch at 2 (diameter 4 mm of the equidistant acanthopore in position, fruit equator, dark 4 mm), inject above-mentioned prescription A1 ~ A3, B1 ~ B3, C1 ~ C3, D1 ~ D3, E1 ~ E3 solution 30 μ L respectively to every hole with micropipettor, CK1 group (sterilized water processed group) and CK2 group (1 * 10 is set simultaneously
8The independent processed group of CFU/ml yeast cream), after 4 hours, inject 1 * 10 to every hole respectively again
4CFU/ml green mold spore suspension 15 μ L, treat that bacterium liquid absorbs after, the single fruit packing is preserved under the environment of 20 ℃ of temperature, relative humidity 85% ~ 95%, carries out the observation of the fruit inoculation incidence of disease and scab diameter respectively at storage on the the 4th, 6,8 day and measures.Handle 5 fruits, triplicate for every group.
The observed result of the citrusfruit green mold inoculation incidence of disease as shown in Figure 1, the incidence of disease minimum of the D2 that as seen fills a prescription group preserve all morbidities of fruit in 6 days, and the incidence of disease of CK1 group is 90%, the incidence of disease that CK2 organizes is 40%; Preserved 8 days, the incidence of disease of prescription D2 group is 20%, and the incidence of disease of CK1 group is 100%, and the incidence of disease of CK2 group is 80%.
The measurement result of citrusfruit green mould spot diameter as shown in Figure 2, the scab diameter minimum of the D2 that as seen fills a prescription group, the scab diameter of preserving 8 days is 59cm, and the scab diameter of CK1 is 106cm, the scab diameter of CK2 is 85cm.
It can also be seen that from Fig. 1 and Fig. 2 the sick effect of control of prescription E2, E3 is also better, it is poor slightly just to compare with prescription D2, and shitosan content and the CaCl of the shitosan content of prescription E2, prescription E3
2The a little higher than prescription of content D2.Therefore, from cost-effectiveness recently, prescription E2, E3 are optimization formula, and prescription D2 is optimum formula.
Three, yeast and chitosan oligosaccharide, shitosan, CaCl
2
Combined Processing is to the influence of the common storage disease of citrusfruit
Prescription D2:0.5% (g/ml) shitosan+1.0% (g/ml) CaCl
2+ 1 * 10
8The CFU/ml yeast cream;
Prescription F1:2.0% (g/ml) chitosan oligosaccharide+0.5% (g/ml) shitosan+1.0% (g/ml) CaCl
2+ 1 * 10
8The CFU/ml yeast cream;
Preparation method: to 1 * 10
8Adding acetic acid to final concentration in the CFU/ml yeast cream is 0.5% (ml/ml), adds a certain amount of chitosan oligosaccharide, shitosan, CaCl again
2To final concentration shown in above-mentioned prescription D2 or the F1, regulate the pH value to 5.8-6, namely with the 0.1mol/L sodium hydroxide solution at last.
Get the surface through the citrusfruit of 75% (ml/ml) alcohol wipe sterilization, be soaked in respectively in above-mentioned prescription D2 and the F1 solution 2 minutes, CK1 group (sterilized water processed group) is set simultaneously, naturally dry back single fruit packing, under the environment of 20 ℃ of temperature, relative humidity 85% ~ 95%, preserve, carried out fruit natural occurrence rate on the the 7th, 14,21,30,40,60 day respectively at storage and observe.Handle 40 fruits, triplicate for every group.
The observed result of citrusfruit natural occurrence rate as shown in Figure 3, the natural occurrence rate of the D2 that as seen fills a prescription group and F1 group is all low than the CK1 group, preserve 7 days, the incidence of disease of CK1 group is 10%, and the fruit that prescription D2 group and F1 organize is not all fallen ill; Preserved 21 days, the incidence of disease of CK1 group is 27.5%, and the incidence of disease of prescription D2 and F1 group is respectively 15% and 7.5%; Preserved 60 days, the incidence of disease of CK1 group is 82.5%, and the incidence of disease of prescription D2 group and F1 group is respectively 60% and 57.5%.Analyze and find, the incidence of disease of prescription D2 group and F1 group has larger difference in earlier stage in storage, but the prolongation along with storage time, difference between two groups is more and more littler, as two groups incidence of disease difference is 5% when preserving 21 days, and two groups incidence of disease difference is decreased to 2.5% when preserving 40 days and 60 days, illustrates that prescription F1 can improve the good fruit rate of storage citrusfruit in early stage, but it is close to preserve the fill a prescription fresh-keeping effect of F1 and D2 of later stage.
The natural occurrence rate observed result of the common storage disease of citrusfruit as shown in Figure 4, wherein scheming A is preceding 20 days each disease incidence of storage period, figure B is each disease incidence of whole storage period (60 days), as seen citrusfruit is in preceding 20 days main pastiness of storage, green mould, fraction is sent out anthracnose and sour rot, and preserves 20 ~ 40 days main blossom-end rot, brown spot and the anthracnose sent out; Compare with prescription D2 group, prescription F1 group has mainly suppressed generation blue or green, green mould in earlier stage in storage, mainly suppressed the generation of anthracnose in the storage later stage, but the incidence of disease at storage later stage blossom-end rot increases, the good fruit rate that causes preserving after 60 days is only organized a little more than prescription D2, therefore, and from cost-effectiveness recently, then the fill a prescription sick better effects if of control of F1 of short-term storage, the Long-term Storage D2 that then fills a prescription has more advantage economically.
Four, yeast and chitosan oligosaccharide, shitosan, CaCl
2
Combined Processing is to the influence of citrusfruit quality
The citrusfruit of surface 75% (ml/ml) alcohol wipe sterilization was soaked in the prescription F1 solution 2 minutes, CK1 group (sterilized water processed group) and CK2 is set simultaneously organizes (1 * 10
8The independent processed group of CFU/ml yeast cream), naturally dry back single fruit packing, under the environment of 20 ℃ of temperature, relative humidity 85% ~ 95%, preserve, carried out fruit weight-loss ratio, soluble solid content, titratable acid and an ascorbic acid (antioxidant) assay in per 7 days.Handle 40 fruits, triplicate for every group.
The measurement result of citrusfruit weight-loss ratio is preserved after 14 days as shown in Figure 5, and the fruit weight-loss ratio of prescription F1 group is lower than CK1 group and CK2 group, and yeast and chitosan oligosaccharide, shitosan, CaCl are described
2Combined Processing more can be slowed down the loss of moisture and water-soluble nutritional labeling in the citrusfruit tissue than the independent processing of yeast.
The assay result of citrusfruit soluble solid preserves after 28 days as shown in Figure 6, and the soluble solid content of prescription F1 group is lower than CK1 group and CK2 group, and yeast and chitosan oligosaccharide, shitosan, CaCl are described
2Combined Processing is better than yeast and handles separately delaying the effect of citrusfruit aspect old and feeble.
The assay result of citrusfruit titratable acid preserves after 21 days as shown in Figure 7, and the titratable acid content of prescription F1 group is higher than CK1 group and CK2 group, and yeast and chitosan oligosaccharide, shitosan, CaCl are described
2Combined Processing is better than yeast and handles separately delaying the effect of citrusfruit aspect old and feeble.
Citrusfruit content of ascorbic acid measurement result is preserved after 7 days as shown in Figure 8, and the ascorbic acid content of prescription F1 group is lower than CK1 group and CK2 group, and yeast and chitosan oligosaccharide, shitosan, CaCl are described
2Combined Processing is better than yeast and handles separately delaying the effect of citrusfruit aspect old and feeble.
In general, prescription F1 can significantly reduce generation and the delayed fruit aging of citrusfruit postharvest disease, and the citrusfruit quality is not had harmful effect; Prescription D2 also has above-mentioned close effect, also has advantage economically when Long-term Storage; F1, D2 are poor slightly though prescription E2, E3 effect are filled a prescription, and can reach the object of the invention too.
Explanation is at last, above embodiment is only unrestricted in order to technical scheme of the present invention to be described, although by invention has been described with reference to the preferred embodiments of the present invention, but those of ordinary skill in the art is to be understood that, can make various changes to it in the form and details, and not depart from the spirit and scope of the present invention that appended claims limits.
Claims (5)
1. the yeast complex biological preservative is characterized in that, contains 1 * 10 among every 100mL
10CFU Pichia membranaefaciens, 0.5g shitosan, 1g calcium chloride and 0.5mL acetic acid, surplus are water, and regulating the pH value is 5.8-6.
2. the yeast complex biological preservative is characterized in that, contains 1 * 10 among every 100mL
10CFU Pichia membranaefaciens, 2g chitosan oligosaccharide, 0.5g shitosan, 1g calcium chloride and 0.5mL acetic acid, surplus are water, and regulating the pH value is 5.8-6.
3. the preparation method of claim 1 or 2 described yeast complex biological preservatives is characterized in that, may further comprise the steps: add acetic acid and water in Pichia membranaefaciens suspension, make every 100mL and contain 1 * 10
10The mixed liquor of CFU yeast and 0.5mL acetic acid adds chitosan oligosaccharide, shitosan and calcium chloride again in this mixed liquor, make among every 100mL and contain 1 * 10
10The mixed liquor of CFU Pichia membranaefaciens, 0g or 2g chitosan oligosaccharide, 0.5g shitosan, 1g calcium chloride and 0.5mL acetic acid, regulating the pH value is 5.8-6, namely gets the yeast complex biological preservative;
Wherein, the preparation method of described Pichia membranaefaciens suspension is: Pichia membranaefaciens is inoculated in the NYDB nutrient solution, and in 28 ± 1 ℃ of shaken cultivation of temperature, the centrifugal supernatant of abandoning, it is resuspended that yeast cells adds water, namely gets Pichia membranaefaciens suspension.
4. claim 1 or 2 application of described yeast complex biological preservative in citrusfruit is fresh-keeping.
5. utilize claim 1 or 2 described yeast complex biological preservatives to carry out the fresh-keeping method of citrusfruit, it is characterized in that, citrusfruit was soaked 2 minutes in the yeast complex biological preservative, namely.
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|---|---|---|---|---|
| CN1302546A (en) * | 2001-03-01 | 2001-07-11 | 中国科学院植物研究所 | Biological antagonistic bacterium (BAY-1) for preventing and treating soft rot of fruit and vegetables and its application method |
| CN1563351A (en) * | 2004-04-21 | 2005-01-12 | 中国科学院植物研究所 | Biologic bactericide for preventing and curing disease of garden of spgarden stuff after picking, and specific strain |
| CN101513211A (en) * | 2009-04-03 | 2009-08-26 | 西南大学 | Complex biological preservative formula of citrus fruit chitosan oligosaccharide |
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2012
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1302546A (en) * | 2001-03-01 | 2001-07-11 | 中国科学院植物研究所 | Biological antagonistic bacterium (BAY-1) for preventing and treating soft rot of fruit and vegetables and its application method |
| CN1563351A (en) * | 2004-04-21 | 2005-01-12 | 中国科学院植物研究所 | Biologic bactericide for preventing and curing disease of garden of spgarden stuff after picking, and specific strain |
| CN101513211A (en) * | 2009-04-03 | 2009-08-26 | 西南大学 | Complex biological preservative formula of citrus fruit chitosan oligosaccharide |
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| Title |
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| 周雅涵 等.拮抗酵母菌对果蔬采后病害生防增效途径及机理研究进展.《食品科学》.2011,第32卷(第17期), |
| 拮抗酵母菌对果蔬采后病害生防增效途径及机理研究进展;周雅涵 等;《食品科学》;20111231;第32卷(第17期);362-365 * |
| 柑橘果实采后侵染性病害防治技术研究进展;邓雨艳 等;《食品科技》;20081231(第4期);211-214 * |
| 邓雨艳 等.柑橘果实采后侵染性病害防治技术研究进展.《食品科技》.2008,(第4期), |
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