CN103999930B - The biological composite mildewproof antistaling agent of a kind of oranges and tangerines and preparation and application - Google Patents
The biological composite mildewproof antistaling agent of a kind of oranges and tangerines and preparation and application Download PDFInfo
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- CN103999930B CN103999930B CN201410242839.6A CN201410242839A CN103999930B CN 103999930 B CN103999930 B CN 103999930B CN 201410242839 A CN201410242839 A CN 201410242839A CN 103999930 B CN103999930 B CN 103999930B
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- 241000675108 Citrus tangerina Species 0.000 title claims abstract description 60
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- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 241000193744 Bacillus amyloliquefaciens Species 0.000 claims abstract description 42
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 41
- 239000012530 fluid Substances 0.000 claims abstract description 39
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 32
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 27
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 24
- 239000012153 distilled water Substances 0.000 claims abstract description 19
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- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims abstract description 4
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- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 10
- 238000004321 preservation Methods 0.000 description 6
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- 102000003992 Peroxidases Human genes 0.000 description 5
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Abstract
The biological composite mildewproof antistaling agent of oranges and tangerines of the present invention is made up of following masses, parts by volume raw material: 1 part, glacial acetic acid, shitosan 2-3 part, bacillus amyloliquefaciens HRH.317 zymotic fluid 1.5-2 part, Tween-80? 1.5-2 part, NaCl? 0.3-0.5 part, distilled water 93 parts.The preparation method of this antistaling agent is: in distilled water, first add glacial acetic acid make acetum; Then in acetum, add shitosan, bacillus amyloliquefaciens HRH317 zymotic fluid, Tween-80 and NaCl respectively, stir.The using method of this antistaling agent is: immerse after not damaged, the cleaning of disease-free oranges and tangerines running water in the liquor natrii hypochloritis of 0.1% 1-2min that sterilizes, the clorox of bearing fruit by sterile water wash after taking-up also dries, then oranges and tangerines are immersed completely this antistaling agent and soak 30-40 second, pull out and dry the packing of rear PE freshness protection package completely, every bag of 25-40 fruit, put into plastic crate be built in the condition of room temperature 18-23 DEG C and indoor relative humidity 70%-80% under or be placed in reserve temperature 4-6 DEG C, preserve under the condition of relative humidity 70%-80%.
Description
Technical field
The present invention relates to oranges and tangerines technical field of preservation of fresh, be specifically related to adopt biological anti-mould fresh keeping agent to carry out preparation and application that is fresh-keeping and anti-mould fresh keeping agent to oranges and tangerines.
Background technology
Fruits and vegetables are one of Major Foods of the mankind, but fruits and vegetables are subject to regional and seasonal impact, easily occur putrid and deteriorated.The putrid and deteriorated meeting of fruits and vegetables causes unnecessary monetary loss, and causes the reduction of fruit-vegetable quality, and wherein food-borne bacterium can produce a large amount of toxin, makes people cause the diseases such as disease of digestive tract, immunological diseases and inflammation.Traditional chemical anti-mildew fresh-keeping liquid is greatly toxic, easily cause the allergy of people and poisoning, and novel biological preservation liquid is only albumen, polypeptide, some lipid and organic acid etc., does not have toxic and side effect or toxic and side effect very little to human body.Biological preservation liquid is the material that a class of direct separation and Extraction from biology has bacteriostasis, can be divided into three types, i.e. animal sources methylene azur, vegetable source natural fresh-keeping liquid and microbial source methylene azur according to source.Therefore, the research and development of new type of safe effective biological fruit and vegetable anti-mildew fresh-keeping liquid are extremely urgent.
Summary of the invention
The object of the invention is for the deficiency of prior art to the research and development of biological preservation liquid, the biological composite mildewproof antistaling agent of a kind of oranges and tangerines and preparation and application are provided.
The biological composite mildewproof antistaling agent of oranges and tangerines of the present invention is made up of following masses, parts by volume raw material: 1 part, glacial acetic acid, shitosan 2-3 part, bacillus amyloliquefaciens (
bacillusamyloliquefaciens) HRH.317 zymotic fluid 1.5-2 part, Tween-80 1.5-2 part, NaCl0.3-0.5 part, distilled water 93 parts.
The preparation method of the biological composite mildewproof antistaling agent of oranges and tangerines of the present invention comprises the steps:
(1) acetum made by the glacial acetic acid adding 1 part (V/V) in 93 parts of distilled water;
(2) add the NaCl of the shitosan of 2-3 part (W/V), the bacillus amyloliquefaciens HRH317 zymotic fluid of 1.5-2 part (V/V), the Tween-80 of 1.5-2 part (W/V) and 0.3-0.5 part (W/V) in the acetum prepared in (1) respectively, stir.
The biological composite mildewproof antistaling agent of oranges and tangerines of the present invention is liquid that is colourless or slightly light yellow clear.
Described bacillus amyloliquefaciens (
bacillusamyloliquefaciens) HRH.317 bacterial strain applied for a patent in the center preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms by Agricultural University Of Shanxi, preserving number is CGMCCJL/0136, number of patent application is 201310187052.X.
The preparation method of described bacillus amyloliquefaciens HRH.317 zymotic fluid comprises the steps:
(1) prepare fluid nutrient medium in 100 parts of distilled water, add 0.5 portion of beef extract, 1 part of peptone and 0.5 part of NaCl, after regulating pH to 7.2, sterilizing in 121 DEG C, 20 minutes, makes nutrient solution for subsequent use;
(2) activation of bacillus amyloliquefaciens HRH317 aseptically, bacillus amyloliquefaciens HRH.317 is inoculated in oese in the nutrient solution (1) prepared, under 37 DEG C of conditions, rotating speed is shaken cultivation 24h in the constant-temperature table of 150r/min, for subsequent use as seed liquor;
(3) by seed liquor that (2) have activated, in the fluid nutrient medium prepared with 1% (V/V) inoculum concentration access (1), be shaken cultivation 24h in the constant-temperature table of 150r/min at rotating speed, obtain bacillus amyloliquefaciens HRH.317 basal fermentation liquid;
(4) in basal fermentation liquid access (1) fluid nutrient medium prepared prepared by (3), 36-38 DEG C, under 160r/min condition, cultivate 20-24h, expand cultivation and obtain zymotic fluid, zymotic fluid viable count is 10
7-10
8cfu/mL.
Bacillus amyloliquefaciens HRH.317 zymotic fluid proves safe through the anxious poison experiment of animal used as test.
The using method of the biological composite mildewproof antistaling agent of oranges and tangerines of the present invention comprises the steps:
(1) by without withered, not damaged, disease-free oranges and tangerines, immerse in the liquor natrii hypochloritis of 0.1% with after running water cleaning the 1-2min that sterilizes, with the clorox that sterile water wash is born fruit after taking-up, and dry;
(2) oranges and tangerines after drying are immersed biological composite mildewproof antistaling agent of the present invention completely, soak 30-40 second, pull out and dry PE (polyethylene) freshness protection package that rear thickness is 0.2-0.4mm completely and (punch at interval of 15cm place on bag in advance, bore dia is 1cm) packing, every bag of 25-40 fruit, stay 1/4 sack space, put into plastic crate be built in the condition of room temperature 18-23 DEG C and indoor relative humidity 70%-80% under or be placed in reserve temperature 4-6 DEG C, preserve under the condition of relative humidity 70%-80%.
The anti-mildew fresh-keeping effect test of the biological composite mildewproof antistaling agent of oranges and tangerines of the present invention
1 test method
1.1 test materials: the oranges and tangerines 400 that crust is intact.
1.2 processing methods: the A intact oranges and tangerines of biological composite mildewproof antistaling agent process crust of the present invention, processing method is with above-mentioned using method; B contrast 1 is the oranges and tangerines that crust is intact; C biological composite mildewproof antistaling agent of the present invention process manually punctures the intact citrus fruit peel inoculation mould spore of crust, and processing method is with above-mentioned using method; D contrast 2 is manually puncture the intact citrus fruit peel inoculation mould spore of crust; Each 100 fruits for the treatment of capacity.
Whether the final conclusion that chemical anti mildew agent is also ununified can be used on oranges and tangerines due to current, simultaneously distributors employs chemical anti mildew agent to the oranges and tangerines that market is sold and all takes to avoid or do not take a stand, therefore, under the prerequisite of not knowing concrete chemical preservation mould dose of kind and working concentration, be difficult to determine a suitable chemical anti mildew agent contrast.
1.3 holding conditions and time: A holding conditions is temperature 4-6 DEG C, relative humidity 70%-80%, 60 days time; B holding conditions is 18-23 DEG C, relative humidity 70%-80%, 20 days time; C holding conditions is temperature 18-21 DEG C, relative humidity 70%-80%, 15 days time.
1.4: the mould variability of oranges and tangerines, peroxidase content, mda content, weight-loss ratio, soluble solid content, titratable acid content, ascorbic acid content.
2 result of the tests
2.1 holding conditions are temperature 4-6 DEG C, relative humidity inspection target 70%-80%, when preserving 60 days, mould variability through anti-mould fresh keeping agent process oranges and tangerines of the present invention is 9%, and the mould variability of oranges and tangerines contrasting 1 is 28%, processes and makes the mould variability of oranges and tangerines reduce 19 percentage points than contrast 1; Holding conditions is 18-23 DEG C, relative humidity 70%-80%, and when preserving 20 days, the mould variability through anti-mould fresh keeping agent process oranges and tangerines of the present invention is 14%, and the mould variability of oranges and tangerines contrasting 1 is 40%, processes and makes the mould variability of oranges and tangerines reduce 26 percentage points than contrast 1.
2.2 holding conditions are temperature 18-21 DEG C, relative humidity 70%-80%, and during 15 days time, the mould variability of oranges and tangerines of C process is 35%, and the mould variability of oranges and tangerines contrasting 2 is 100%, process and make the mould variability of oranges and tangerines reduce 65 percentage points than contrast 2.And process the peroxidase content 12.2 Δ OD of C oranges and tangerines
470/ g.Fw.min, mda content 21.5mmol/gFw, weight-loss ratio 2.2%, soluble solid content 9.6%, titratable acid content 0.61%, ascorbic acid content 13.7mg/100g; And contrast the peroxidase content 7.7 Δ OD of 2 oranges and tangerines
470/ g.Fw.min, mda content 25.4mmol/gFw, weight-loss ratio 4.1%, soluble solid content 8.8%, titratable acid content 0.53%, ascorbic acid content 11.6mg/100g.Wherein, the peroxidase of high-load represents the high resistance to cold and diseases of oranges and tangerines, and it is low that the MDA of low content represents oranges and tangerines degree of spoilage, and oranges and tangerines can preserve the longer time; The weight-loss ratio of low value, it is fresher that the soluble solid of high level, titratable acid are anti-, ascorbic acid content represents oranges and tangerines.
As can be seen here, the anti-mildew fresh-keeping of the biological composite mildewproof antistaling agent of oranges and tangerines of the present invention has extremely significant effect.
beneficial effect of the present invention
Above-mentioned result of the test shows, use oranges and tangerines of the present invention biological composite mildewproof antistaling agent process oranges and tangerines, significantly can reduce the mould variability of oranges and tangerines in storage, the peroxidase content of fruit, soluble solid content, titratable acid content and ascorbic acid content can also be improved simultaneously, and mda content and weight-loss ratio can be reduced, strengthen the resistance of fruit, keep the freshness of fruit, reduce the wastage, extend storage period.And avoid the harm of chemical preservative, improve edible security.
Detailed description of the invention
embodiment 1
The biological composite mildewproof antistaling agent of oranges and tangerines of the present invention prepared by employing following masses, parts by volume raw material: 1 part, glacial acetic acid, shitosan 2 parts, bacillus amyloliquefaciens (
bacillusamyloliquefaciens) HRH.317 zymotic fluid 1.5 parts, Tween-80 1.5 parts, NaCl0.3 part, distilled water 93 parts.
The preparation method of bacillus amyloliquefaciens HRH.317 zymotic fluid is:
(1) prepare fluid nutrient medium in 100 parts of distilled water, add 0.5 portion of beef extract, 1 part of peptone and 0.5 part of NaCl, after regulating pH to 7.2, sterilizing in 121 DEG C, 20 minutes, makes nutrient solution;
(2) activation of bacillus amyloliquefaciens HRH317 aseptically, bacillus amyloliquefaciens HRH.317 is inoculated in oese in the nutrient solution (1) prepared, under 37 DEG C of conditions, rotating speed is shaken cultivation 24h in the constant-temperature table of 150r/min, as seed liquor;
(3) by seed liquor that (2) have activated, in the fluid nutrient medium prepared with 1% (V/V) inoculum concentration access (1), be shaken cultivation 24h in the constant-temperature table of 150r/min at rotating speed, obtain bacillus amyloliquefaciens HRH.317 basal fermentation liquid;
(4) in basal fermentation liquid access (1) fluid nutrient medium prepared prepared by (3), 36-38 DEG C, under 160r/min condition, cultivate 20-24h, expand cultivation and obtain zymotic fluid, zymotic fluid viable count can reach 10
7-10
8cfu/mL.
The preparation method of the biological composite mildewproof antistaling agent of oranges and tangerines is: acetum made by the glacial acetic acid adding 1 part (V/V) in 93 parts of distilled water; In acetum, add the shitosan of 2 parts (W/V), the bacillus amyloliquefaciens HRH317 zymotic fluid of 1.5 parts (V/V), the Tween-80 of 1.5 parts (W/V) and the NaCl of 0.3 part (W/V) respectively, stir.
The using method of the biological composite mildewproof antistaling agent of oranges and tangerines comprises the steps:
(1) by without withered, not damaged, disease-free oranges and tangerines, immerse in the liquor natrii hypochloritis of 0.1% with after running water cleaning the 1-2min that sterilizes, with the clorox that sterile water wash is born fruit after taking-up, and dry;
(2) oranges and tangerines after drying are immersed biological composite mildewproof antistaling agent of the present invention completely, soak 30-40 second, pull out and dry PE (polyethylene) freshness protection package that rear thickness is 0.2-0.4mm completely and (punch at interval of 15cm place on bag in advance, bore dia is 1cm) packing, every bag of 25-40 fruit, stay 1/4 sack space, put into plastic crate be built in the condition of room temperature 18-23 DEG C and indoor relative humidity 70%-80% under or be placed in reserve temperature 4-6 DEG C, preserve under the condition of relative humidity 70%-80%.
embodiment 2
The biological composite mildewproof antistaling agent of oranges and tangerines of the present invention prepared by employing following masses, parts by volume raw material: 1 part, glacial acetic acid, shitosan 3 parts, bacillus amyloliquefaciens (
bacillusamyloliquefaciens) HRH.317 zymotic fluid 2 parts, Tween-80 2 parts, NaCl0.5 part, distilled water 93 parts.
The preparation method of bacillus amyloliquefaciens HRH.317 zymotic fluid is with embodiment 1.
The preparation method of the biological composite mildewproof antistaling agent of oranges and tangerines is: acetum made by the glacial acetic acid adding 1 part (V/V) in 93 parts of distilled water; In acetum, add the shitosan of 3 parts (W/V), the bacillus amyloliquefaciens HRH317 zymotic fluid of 2 parts (V/V), the Tween-80 of 2 parts (W/V) and the NaCl of 0.5 part (W/V) respectively, stir.
The using method of the biological composite mildewproof antistaling agent of oranges and tangerines is with embodiment 1.
embodiment 3
The biological composite mildewproof antistaling agent of oranges and tangerines of the present invention prepared by employing following masses, parts by volume raw material: 1 part, glacial acetic acid, shitosan 2.5 parts, bacillus amyloliquefaciens (
bacillusamyloliquefaciens) HRH.317 zymotic fluid 1.8 parts, Tween-80 1.7 parts, NaCl0.4 part, distilled water 93 parts.
The preparation method of bacillus amyloliquefaciens HRH.317 zymotic fluid is with embodiment 1.
The preparation method of the biological composite mildewproof antistaling agent of oranges and tangerines is: acetum made by the glacial acetic acid adding 1 part (V/V) in 93 parts of distilled water; In acetum, add the shitosan of 2.5 parts (W/V), the bacillus amyloliquefaciens HRH317 zymotic fluid of 1.8 (V/V), the Tween-80 of 1.7 parts (W/V) and the NaCl of 0.4 part (W/V) respectively, stir.
embodiment 4
The biological composite mildewproof antistaling agent of oranges and tangerines of the present invention prepared by employing following masses, parts by volume raw material: 1 part, glacial acetic acid, shitosan 2 parts, bacillus amyloliquefaciens (
bacillusamyloliquefaciens) HRH.317 zymotic fluid 1.5 parts, Tween-80 1.5 parts, NaCl0.3 part, distilled water 93 parts.
The preparation method of bacillus amyloliquefaciens HRH.317 zymotic fluid is with embodiment 1.
The preparation method of the biological composite mildewproof antistaling agent of oranges and tangerines is: acetum made by the glacial acetic acid adding 1 part (V/V) in 93 parts of distilled water; In acetum, add the shitosan of 2 parts (W/V), the bacillus amyloliquefaciens HRH317 zymotic fluid of 1.5 parts (V/V), the Tween-80 of 1.5 parts (W/V) and the NaCl of 0.3 part (W/V) respectively, stir.
embodiment 5
The biological composite mildewproof antistaling agent of oranges and tangerines of the present invention prepared by employing following masses, parts by volume raw material: 1 part, glacial acetic acid, shitosan 3 parts, bacillus amyloliquefaciens (
bacillusamyloliquefaciens) HRH.317 zymotic fluid 2 parts, Tween-80 2 parts, NaCl0.5 part, distilled water 93 parts.
The preparation method of bacillus amyloliquefaciens HRH.317 zymotic fluid is with embodiment 1.
The preparation method of the biological composite mildewproof antistaling agent of oranges and tangerines is: acetum made by the glacial acetic acid adding 1 part (V/V) in 93 parts of distilled water; In acetum, add the shitosan of 3 parts (W/V), the bacillus amyloliquefaciens HRH317 zymotic fluid of 2 (V/V), the Tween-80 of 2 parts (W/V) and the NaCl of 0.5 part (W/V) respectively, stir.
Claims (2)
1. the biological composite mildewproof antistaling agent of oranges and tangerines, be made up of following masses, parts by volume raw material: 1 part, glacial acetic acid, shitosan 2-3 part, bacillus amyloliquefaciens (Bacillusamyloliquefaciens) HRH.317 zymotic fluid 1.5-2 part, Tween-80 1.5-2 part, NaCl0.3-0.5 part, distilled water 93 parts;
The preparation method of the biological composite mildewproof antistaling agent of described oranges and tangerines comprises the steps:
(1) acetum made by the glacial acetic acid adding 1 part (V/V) in 93 parts of distilled water;
(2) add the NaCl of the shitosan of 2-3 part (W/V), the bacillus amyloliquefaciens HRH317 zymotic fluid of 1.5-2 part (V/V), the Tween-80 of 1.5-2 part (W/V) and 0.3-0.5 part (W/V) in the acetum prepared in step (1) respectively, stir;
It is characterized in that, the preparation method of described bacillus amyloliquefaciens HRH.317 zymotic fluid comprises the steps:
A prepares fluid nutrient medium: in 100 parts of distilled water, add 0.5 portion of beef extract, 1 part of peptone and 0.5 part of NaCl, and after regulating pH to 7.2, sterilizing in 121 DEG C, 20 minutes, makes nutrient solution;
The activation of B bacillus amyloliquefaciens HRH317: aseptically, bacillus amyloliquefaciens HRH.317 is inoculated in nutrient solution prepared by steps A with oese, under 37 DEG C of conditions, rotating speed is shaken cultivation 24h in the constant-temperature table of 150r/min, as seed liquor;
The seed liquor that step B activates by C, in the fluid nutrient medium prepared with 1% (V/V) inoculum concentration access steps A, is shaken cultivation 24h in the constant-temperature table of 150r/min at rotating speed, obtains bacillus amyloliquefaciens HRH.317 basal fermentation liquid;
In fluid nutrient medium prepared by basal fermentation liquid that step C is prepared by D access steps A, 36-38 DEG C, under 160r/min condition, cultivate 20-24h, expand cultivation and obtain zymotic fluid, zymotic fluid viable count is 107-108cfu/mL.
2. the biological composite mildewproof antistaling agent of oranges and tangerines according to claim 1, is characterized in that the using method of the biological composite mildewproof antistaling agent of oranges and tangerines is:
(1) by without withered, not damaged, disease-free oranges and tangerines, immerse in the liquor natrii hypochloritis of 0.1% with after running water cleaning the 1-2min that sterilizes, with the clorox that sterile water wash is born fruit after taking-up, and dry;
(2) oranges and tangerines after drying are immersed composite mildewproof antistaling agent completely, soak 30-40 second, pull out and dry the PE freshness protection package packing that rear thickness is 0.2-0.4mm completely, every bag of 25-40 fruit, stay 1/4 sack space, put into plastic crate be built in the condition of room temperature 18-23 DEG C and indoor relative humidity 70%-80% under or be placed in reserve temperature 4-6 DEG C, preserve under the condition of relative humidity 70%-80%.
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|---|---|---|---|
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|---|---|---|---|---|
| CN104430834B (en) * | 2014-12-30 | 2017-10-20 | 华南师范大学 | Applications of the Bacillus amyloliquefaciens strain B014 in postharvest fruit and vegetable is fresh-keeping |
| CN106720241A (en) * | 2016-11-17 | 2017-05-31 | 天津捷盛东辉保鲜科技有限公司 | Grape biology anti-mildew fresh-keeping method |
| CN106615085B (en) * | 2016-12-19 | 2020-01-10 | 山西省农业科学院生物技术研究中心 | Microbial and plant source composite fruit and vegetable fresh-keeping agent |
| CN113412858A (en) * | 2021-04-28 | 2021-09-21 | 四川大学 | Method for preserving blood orange by using bacillus amyloliquefaciens M73 fermentation product |
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| CN102191200A (en) * | 2011-03-31 | 2011-09-21 | 国家粮食局科学研究院 | Bacillus amyloliquefaciens and application thereof |
| CN102524910A (en) * | 2012-01-10 | 2012-07-04 | 中国科学院烟台海岸带研究所 | Edible food fresh-keeping coating and preparation method thereof |
| CN103333828A (en) * | 2013-06-27 | 2013-10-02 | 中国农业大学 | Application of anti-mildew bacillus strain and antimicrobial product thereof in corn storage |
| CN103572597A (en) * | 2013-10-24 | 2014-02-12 | 浙江理工大学 | Antibacterial reinforcement method of rotten cotton fabric cultural relics |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102191200A (en) * | 2011-03-31 | 2011-09-21 | 国家粮食局科学研究院 | Bacillus amyloliquefaciens and application thereof |
| CN102524910A (en) * | 2012-01-10 | 2012-07-04 | 中国科学院烟台海岸带研究所 | Edible food fresh-keeping coating and preparation method thereof |
| CN103333828A (en) * | 2013-06-27 | 2013-10-02 | 中国农业大学 | Application of anti-mildew bacillus strain and antimicrobial product thereof in corn storage |
| CN103572597A (en) * | 2013-10-24 | 2014-02-12 | 浙江理工大学 | Antibacterial reinforcement method of rotten cotton fabric cultural relics |
Non-Patent Citations (1)
| Title |
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| 柑橘绿霉病拮抗细菌的筛选及其抑制效果;安国栋等;《华中农业大学学报》;20130531;第32卷(第3期);第52页摘要 * |
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