CN104316478A - Determination method of hypsizigus marmoreus fungi pack ripeness - Google Patents
Determination method of hypsizigus marmoreus fungi pack ripeness Download PDFInfo
- Publication number
- CN104316478A CN104316478A CN201410583701.2A CN201410583701A CN104316478A CN 104316478 A CN104316478 A CN 104316478A CN 201410583701 A CN201410583701 A CN 201410583701A CN 104316478 A CN104316478 A CN 104316478A
- Authority
- CN
- China
- Prior art keywords
- bacterium bag
- ripeness
- sample
- gill fungus
- spot
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Abstract
The invention relates to a set of determination method of hypsizigus marmoreus fungi pack ripeness. The appearance index, physical and chemical index and biological parameters in the cultivation material according to different cultivation days are determined through the method, the water content, total sugar content, reducing sugar content, and soluble protein content of the fungi pack and the enzyme activity in the fungi pack are determined so as to determine the ripeness of the hypsizigus marmoreus fungi pack. The determination method is capable of providing evidence for the determination of the fungi pack ripeness in the production process of the hypsizigus marmoreus; and meanwhile, the establishment of various inner biological parameters and biochemical index provides new perspective for the research on the after-ripening research theory in edible fungi industry.
Description
Technical field
The invention belongs to biology fungi field, relate to the decision method of a set of spot beautiful gill fungus bacterium bag degree of ripeness specifically.
Background technology
Spot beautiful gill fungus Hypsizygus marmoreus (Peck) H.E. Bigelow has another name called beautiful gill fungus, belong to Agaricales Agaricales, Tricholomataceae Tricholomataceae, beautiful gill fungus belongs to Hypsizygus, it is a kind of low in calories, low-fat health food, is just causing more and more concern focusing on the people of healthy diet.And its taste is fresher than flat mushroom, meat is thicker than sliding mushroom, and matter is more tough than mushroom, excellent taste, also has unique crab fragrance, has saying of " fragrant at matsutake, taste beautiful gill fungus ", therefore have again true Ji mushroom one title in Japan.Listed in menu by more and more fashionable restaurant.
The beautiful gill fungus of spot is one of important kind of Fujian Province's factorial praluction in recent years.Current Fujian is the beautiful gill fungus production base of national important spot, and output accounts for national about 60-70%.The beautiful gill fungus of spot is a fruiting performances at low temperature kind, conventional cultivation is subject to the impact of envirment factor, causes output unstable, with low quality, commodity value is low, because it sends out bacterium comparatively slowly in cultivation is produced, the After-mature cultivation time is long, and the production cycle reaches more than 120-150 days, and be subject to outside environmental elements impact, therefore, there is many problems in process of production, current spot beautiful gill fungus batch production spot is beautiful, and gill fungus is common there is following problem:
(1) different manufacturers bacterium packaging doses differs, and output height is different, and biological efficiency cannot be sought unity of standard, and utilization rate of raw materials is low, and production cost is uncontrollable;
(2) to cultivate degree of ripeness general by rule of thumb for bacterium bag, and using bacterium bag incubation time and mode of appearance as bacterium bag degree of ripeness basis for estimation, cause every batch of fruiting irregular, raw material utilizes insufficient, even occurs thick mycoderma, causes not fruiting phenomenon, brings about great losses;
(3) because winter temperature is low, mycelial growth is slow, although incubation time is enough, often not, cause and yield poorly, raw material availability is low, traces it to its cause and mainly cannot accurately judge bacterium bag degree of ripeness for bacterium bag feed degradation and nutrient accumulation.
Therefore, set up the key technical problem that the inherent biological parameter of judgement bacterium bag degree of ripeness of science and biochemical indicator are the beautiful gill fungus standardized production of spot, this has important practical significance to the beautiful gill fungus factorial praluction of efficient stable spot.
The beautiful gill fungus of spot is one of important kind of Fujian Province's factorial praluction in recent years.Current Fujian is the beautiful gill fungus production base of national important spot, and output accounts for national about 60-70%.But the beautiful gill fungus of spot is cultivation phase longer edible fungi, and approximately need 120-150 days, this through a physiological maturity (latter stage of ripening), just must can grow good mushroom flower bud mainly due to the beautiful gill fungus cultural hypha of spot.Therefore, the mensuration of spot beautiful gill fungus cultural hypha process maturity model directly affects biological efficiency and the output of the beautiful gill fungus of spot.
?
Summary of the invention
The present invention mainly provides the assay method of a set of spot beautiful gill fungus bacterium bag degree of ripeness, the method by measure spot beautiful gill fungus bacterium bag appearance index, physical and chemical index and biological parameter can distinguish easily which bacterium bag maturation can carry out fruiting, which bacterium bag does not also reach physiological maturity can not fruiting.
For achieving the above object, the present invention adopts following technical scheme:
1. the beautiful gill fungus bacterium of spot is bundled into ripe being outside one's consideration and sees the foundation of judge index
(1) different incubation time mycelia color and metamorphosis degree are observed;
(2) the different incubation time of bacterium bag and the flexible mutation analysis of bacterium bag;
2. the foundation of spot beautiful gill fungus bacterium bag degree of ripeness physical and chemical index
(1) in the beautiful gill fungus planting material of spot, C/N measures containing quantitative analysis;
(2) bacterium bag incubation pH, change of moisture content and the relationship analysis of bacterium bag degree of ripeness;
(3) bacterium bag incubation weight change and the relationship analysis of bacterium bag degree of ripeness;
(4) bacterium bag incubation nutrient accumulation and bacterium bag degree of ripeness correlation analysis.
3. the foundation of spot beautiful gill fungus bacterium bag degree of ripeness biological parameter
(1) analysis on change of bacterium bag incubation key enzyme-cellulase activity;
(2) analysis on change of bacterium bag incubation key enzyme-proteinase activity;
(3) analysis on change of bacterium bag incubation key enzyme-xylanase activity;
(4) analysis on change of bacterium bag incubation key enzyme-amylase activity;
(5) analysis on change of bacterium bag incubation key enzyme-lignin oxidation's enzymatic activity;
(6) analysis on change of bacterium bag incubation key enzyme-manganese peroxidase enzymatic activity;
(5) analysis on change of bacterium bag incubation key enzyme-laccase activity;
4. the research of bacterium bag degree of ripeness and biological efficiency relation
(1) the bacterium bag of differing maturity and the beautiful gill fungus Relationship with Yield of spot are analyzed;
(2) the bacterium bag of differing maturity and the research of Fruiting quality relation.
By the foundation of four Iarge-scale system, carry out Accurate Measurement from the degree of ripeness of multiple dimension to the beautiful gill fungus of spot.
Concrete grammar is as follows:
An assay method for spot beautiful gill fungus bacterium bag degree of ripeness, by the process of spot beautiful gill fungus bacterium sample packet, mensuration bacterium comprises the enzyme activity in the water yield, total sugar content, content of reducing sugar, soluble protein content and bacterium bag, measures spot beautiful gill fungus bacterium bag degree of ripeness.
Described sample handling procedure is: spot beautiful gill fungus bacterium bag is cultivated and started in 140 days for 60 days, got every 10 days weight difference within 2g and in 10 days weight reduce bacterium bag at the bacterium bag of 7g-9g as laboratory sample; Bacterium bag is divided into upper, middle and lower three part every 6cm, respectively compost is stirred, accurately take 10 g composts in 50 mL beakers, add 40 mL distilled water and shake up 2 h in 25 DEG C of shaking tables, get liquid in 50 mL centrifuge tubes, centrifugal 15 min of 9000 r/min, supernatant is crude enzyme liquid; Separately take 20 g in plate, dry to constant weight for 80 DEG C, measure water cut.
Described
total sugar content assay method: accurately take the planting material of 0.25 g oven dry to constant weight in 10 mL centrifuge tubes, add 5 mL distilled waters and 1.5 mL concentrated hydrochloric acids, 3 h are hydrolyzed in boiling water bath, supernatant is poured into 50 mL conical flasks after leaving standstill cooling, rejoin 5 mL distilled waters and 1.5 mL concentrated hydrochloric acids, be hydrolyzed 2 h in boiling water bath, after leaving standstill cooling, supernatant be incorporated in 50 mL conical flasks, be settled to 50 mL with distilled water washing.Get 1 mL after mixing and be again settled to 50 mL.Then centrifugal 15 min of 9000 r/min.Get 1 mL supernatant, add 1 mL 5 % phenol, add the 5 mL concentrated sulphuric acids fast, leave standstill 10 min, mixing, 30 water-bath 20 min.490 nm measure absorbance.Sample absorbance substitutes into the total sugar concentration of typical curve regression equation and derived sample.Total sugar concentration × Sample Dilution the multiple of sample total sugar concentration μ g/mL=Sample Dilution.
Described
moisture determination method: after using calibration, pH meter measures supernatant and measures pH, and record plate original weight, weighs plate every day and sample general assembly (TW) no longer reduces to weight, illustrates and dries completely, and record data also calculate water cut.
Described
content of reducing sugar assay method: accurately take planting material 0.5 g of oven dry to constant weight in 10 mL centrifuge tubes, add 5 mL distilled waters in boiling water bath, boil 2 h, then centrifugal 15 min of 9000 r/min, get 1 mL supernatant, add 1.5 mL DNS reagent, boiling water bath 7 min again, then use cold flow water cooling cessation reaction, be settled to 10 mL with distilled water, fully mix, under 540 nm, survey absorbance, and substitute into glucose standard curve regression equation and find corresponding glucose content.
Described
soluble protein content assay method: get 1 mL supernatant, add the Coomassie brilliant G-250 working fluid of 5 mL, develop the color after 10 min, ultraviolet spectrophotometer measures 595 nm place light absorption values, distilled water is as blank, sample absorbance substitutes into the protein concentration that bovine serum albumin(BSA) typical curve regression equation tries to achieve sample, the protein concentration × Sample Dilution multiple of sample protein matter concentration μ g/ mL=Sample Dilution.
Described
enzyme activity determination method in bacterium bag: under 520 nm, survey absorbance, and find corresponding glucose content from glucose standard curve, be calculated to be enzyme activity; Enzyme activity unit is defined as: the hydrolysis substrate per minute enzyme amount generated needed for 1 μ g glucose is defined as 1 enzyme activity unit " U " under these conditions.
The invention has the advantages that: Ban Yu gill fungus manufacturing enterprise mainly relies on bacterium bag incubation time and experience to measure the degree of ripeness of bacterium bag at present, because Ban Yu gill fungus manufacturing enterprise exists larger difference in planting material composition, charge, condition of culture and way to manage, cause bacterium bag degree of ripeness and bacterium bag incubation time often inconsistent, enterprise because of cannot accurately judge bacterium bag whether maturation cause that biological efficiency is low and output is not high, sometimes even there is thick mycoderma not fruiting phenomenon, cause huge economic loss.The method is simple, accurately, can be spot beautiful gill fungus factory culture process bacterium bag degree of ripeness and provides a science judgment foundation.
Embodiment
embodiment 1
sampling and sample preparationspot beautiful gill fungus bacterium bag is cultivated and is started in 140 days for 60 days, gets weight difference every 10 days within 2g, and in 10 days, weight reduces bacterium bag at the bacterium bag of 7g-9g as laboratory sample.Bacterium bag is divided into upper, middle and lower three part every 6cm.Respectively stirred by compost, accurately take 10 g composts in 50 mL beakers, add 40 mL distilled water and shake up 2 h in 25 DEG C of shaking tables, get liquid in 50 mL centrifuge tubes, centrifugal 15 min of 9000 r/min, supernatant is crude enzyme liquid.Separately take 20 g in plate, dry to constant weight for 80 DEG C, measure water cut.
1. the beautiful gill fungus bacterium of spot is bundled into ripe being outside one's consideration and sees the foundation of judge index
Postvaccinal spot beautiful gill fungus bacterium bag 25 DEG C of constant incubators are cultivated, and timing was taken pictures and measured Hyphal length every day, and recorded the distance of mycelia its colour changed into yellow length and bacterium bag deliquescing partial distance sack.Thus determine that difference is observed and the flexible mutation analysis of bacterium bag with incubation time mycelia color and metamorphosis degree.
2. the foundation of spot beautiful gill fungus bacterium bag degree of ripeness physical and chemical index
(1) in the beautiful gill fungus planting material of spot, C/N measures containing quantitative analysis
total sugar content measuresaccurately take the planting material of 0.25 g oven dry to constant weight in 10 mL centrifuge tubes, add 5 mL distilled waters and 1.5 mL concentrated hydrochloric acids, 3 h are hydrolyzed in boiling water bath, supernatant is poured into 50 mL conical flasks after leaving standstill cooling, rejoin 5 mL distilled waters and 1.5 mL concentrated hydrochloric acids, be hydrolyzed 2 h in boiling water bath, after leaving standstill cooling, supernatant be incorporated in 50 mL conical flasks, be settled to 50 mL with distilled water washing.Get 1 mL after mixing and be again settled to 50 mL.Then centrifugal 15 min of 9000 r/min.Get 1 mL supernatant, add 1 mL 5 % phenol, add the 5 mL concentrated sulphuric acids fast, leave standstill 10 min, mixing, 30 water-bath 20 min.490 nm measure absorbance.Sample absorbance substitutes into the total sugar concentration of typical curve regression equation and derived sample.Total sugar concentration × Sample Dilution the multiple of sample total sugar concentration (μ g/mL)=Sample Dilution.
(2) bacterium bag incubation pH, change of moisture content and the relationship analysis of bacterium bag degree of ripeness
After using calibration, pH meter measures supernatant and measures pH.Record plate original weight, weighs plate every day and sample general assembly (TW) no longer reduces to weight, illustrates and dries completely, and record data also calculate water cut.
(3) bacterium bag incubation weight change and the relationship analysis of bacterium bag degree of ripeness
Get postvaccinal spot beautiful gill fungus bacterium bag 20 and be wrapped in 25 DEG C of constant temperature culture rooms cultivations, weighed every 5 days and record a weight.
(4) bacterium bag incubation nutrient accumulation and bacterium bag degree of ripeness correlation analysis.
content of reducing sugar measuresaccurately take planting material 0.5 g of oven dry to constant weight in 10 mL centrifuge tubes, add 5 mL distilled waters in boiling water bath, boil 2 h.Then centrifugal 15 min of 9000 r/min.Get 1 mL supernatant, add 1.5 mL DNS reagent, then boiling water bath 7 min, then use cold flow water cooling cessation reaction, be settled to 10 mL with distilled water, fully mix.Under 540 nm, survey absorbance, and substitute into glucose standard curve regression equation and find corresponding glucose content.
soluble protein measuresget 1 mL supernatant, add the Coomassie brilliant G-250 working fluid of 5 mL, after 10 min that develop the color, ultraviolet spectrophotometer measures 595 nm place light absorption values.Distilled water is as blank.Sample absorbance substitutes into the protein concentration that bovine serum albumin(BSA) typical curve regression equation tries to achieve sample.Protein concentration × Sample Dilution the multiple of sample protein matter concentration (μ g/ mL)=Sample Dilution.
3. the foundation of spot beautiful gill fungus bacterium bag degree of ripeness biological parameter
(1) analysis on change of bacterium bag incubation key enzyme-cellulase activity;
In test tube, add 1 mL 1% sodium carboxymethyl cellulose, after 50 DEG C of preheating 5 min, add 0.5 mL crude enzyme liquid, 50 DEG C of insulation 60 min, add 1.5 mL DNS reagent in test tube, then boiling water bath 7 min, then use cold flow water cooling cessation reaction, be settled to 10 mL with distilled water, fully mix.Blank is done with inactivator liquid.Under 520 nm, survey absorbance, and find corresponding glucose content from glucose standard curve, be calculated to be enzyme activity.Enzyme activity unit is defined as: the hydrolysis substrate per minute enzyme amount generated needed for 1 μ g glucose is defined as 1 enzyme activity unit " U " under these conditions.
(2) analysis on change of bacterium bag incubation key enzyme-proteinase activity
Get 2 mL centrifuge tubes, add crude enzyme liquid liquid 0.5 mL, be placed in 40 DEG C of water-bath preheating 2 min, respectively add casein 0.5 mL through same preheating again, accurately after insulation 30 min, add 0.4 mol/L trichloroacetic acid 1 mL immediately, with cessation reaction, centrifugal 2 min of 9000 r/min, then separately get 10 mL centrifuge tubes, add supernatant 1 mL, add 0.4 mol/L sodium carbonate 5 mL again, add Folin reagent 100 μ L, shake up, after 40 DEG C of insulation color development 20 min, carry out spectrodensitometry.Control test assay method is the same, before adding casein, only first adds 0.4 mol trichloroacetic acid 1 mL, add casein again after making enzyme deactivation.Absorbance substitutes into TYR typical curve regression equation can try to achieve generation TYR amount, is calculated to be enzyme activity.Enzyme activity unit is defined as: at 40 DEG C, caseinhydrolysate per minute produces 1 μ g TYR, is defined as 1 protease activity unit of force " U ".
(3) analysis on change of bacterium bag incubation key enzyme-xylanase activity
Draw 1% xylan 0.75 mL in test tube, after 50 DEG C of preheating 5 min, add 0.25 mL crude enzyme liquid, 50 DEG C of insulation 30 min, add 1.5 mL DNS reagent in test tube, then boiling water bath 7 min, then use cold flow water cooling cessation reaction, be settled to 10 mL with distilled water, fully mix.Blank is done with inactivator liquid.Absorbance is surveyed under 550 nm.Enzyme activity unit is defined as: 50 DEG C, hydrolysis substrate per minute generates needed for 1 μ g D-wood sugar under the condition of pH5.2 enzyme amount is defined as 1 enzyme activity unit " U ".
(4) analysis on change of bacterium bag incubation key enzyme-amylase activity
Draw 4 mL soluble starch solution in test tube, add phosphate buffer 1 mL, after shaking up in 50 DEG C of waters bath with thermostatic control preheating 5min.Add crude enzyme liquid 500 ul, shake up timing, in 50 DEG C of reaction 30 min.Draw reactant liquor 1 mL immediately, add in the test tube filling 0.5 mL watery hydrochloric acid and the rare iodine liquid of 5 mL, shake up.And with 0.5 mL watery hydrochloric acid and 5 mL iodine dilutions for blank, under 660 nm wavelength, measure light absorption value.According to regression equation, calculate the concentration of starch in the rear solution of reaction.Amylase activity unit definition: at 60 DEG C, under pH=6.0 condition, liquefaction 1 μ g soluble starch per minute, is defined as 1 diastatic activity unit of force " U ".
(5) analysis on change of bacterium bag incubation key enzyme-lignin oxidation's enzymatic activity
Accurately pipette HAc-NaAc damping fluid (pH=4.5) 2.7 mL, 1 mmol/L ABTS solution 0.2 mL, mixing shakes up, water-bath preheating 5 min in 30 DEG C of thermostat water baths.Above-mentioned mix reagent being poured into volume is in the cuvette of 3 mL, the zeroing of 420 nm places.Accurately add enzyme liquid 100 μ L to be measured, immediate record light absorption value, records 1 time every 30 s, records 3 min altogether.Enzyme activity unit is defined as: the enzyme amount required for oxidation per minute 1 μm of ol substrate is a Ge Meihuo unit " U ".
(6) analysis on change of bacterium bag incubation key enzyme-manganese peroxidase enzymatic activity
Accurately pipette guaiacol 0.7 mL of 4 mmol/L, 100 mmol/L pH4.5 Na-succinate buffer 2 mL, the H of 2.5 mmol/L
2o
2the MnSO of 5 mmol/L
40.2 mL, mixing, shakes up, water-bath preheating 5 min in 40 DEG C of thermostat water baths.Above-mentioned mix reagent being poured into volume is in the cuvette of 3 mL, the zeroing of 465 nm places.Add 0.2 mL enzyme liquid, immediate record light absorption value, records 1 time every 30 s, records 3 min altogether.Enzyme activity unit is defined as: the enzyme amount of oxidation per minute 1 μm of ol MnSO4 is a Ge Meihuo unit " U ".
(5) analysis on change of bacterium bag incubation key enzyme-laccase activity
Accurately pipette 0.2 mol/L sodium tartrate damping fluid (pH3.0) 1.5 mL, 15 mmol/L veratryl alcohol 1.0 mL, 0.04 mL enzyme liquid, mixing, shakes up, water-bath preheating 5 min in 30 DEG C of thermostat water baths.Above-mentioned mix reagent being poured into volume is in the cuvette of 3 mL, the zeroing of 310 nm places.With 15 mmol/L H
2o
20.01 mL starts reaction, and immediate record light absorption value, records 1 time every 6 s, records 1 min altogether.Enzyme activity unit is defined as: 1 μm of ol veratryl alcohol enzyme amount be oxidized to required for veratraldehyde that makes per minute is a Ge Meihuo unit " U ".
4. the research of bacterium bag degree of ripeness and biological efficiency relation
(1) the bacterium bag of differing maturity and the beautiful gill fungus Relationship with Yield of spot are analyzed
Get each 24 bags of spot beautiful gill fungus bacterium bag that 25 DEG C are cultivated 60,70 to 140 days, weigh and original weight before recording fruiting, carry out management of producing mushroom in 14 DEG C, after plucking, accurate weighing cuts the beautiful gill fungus weight of mushroom pin spot, carries out output significance analysis, weigh residue bacterium bag weight, calculation biology efficiency.
(2) the bacterium bag of differing maturity and the research of Fruiting quality relation
Take pictures and measure spot beautiful gill fungus mushroom handle length with ruler, fructification size thickness, record fructification quality soft or hard degree, get the beautiful gill fungus fructification of 100 g spots and dry to constant weight in 80 DEG C of drying bakers, measure water cut.
According to the appearance index cultivated in different number of days planting material, physical and chemical index and biological parameter measure, obtain when cultivating by the 120th day, bacterium bag upper, middle and lower all becomes light yellow, and it is soft, total sugar content drops to 777.9 μ g/mL and change is mild, upper bottom pH value reaches consistent, bacterium comprises the water yield and reaches 68.7%, content of reducing sugar drops to 516.5 μ g/mL, soluble protein drops to 132.6 μ g/mL, cellulase activity maintains 1.58U/mL, prolease activity drops to 4.02U/mL, xylanase activity rises to 14.1 U/mL, amylase activity declines and maintains about 1.79U/mL, lignin peroxidase vigor rises to 2.41U/mL, manganese peroxidase top is incremented to 40.36U/mL gradually to bottom, the upper bottom of laccase activity all reaches 173.8 U/L, at this moment fruiting output and biological efficiency the highest.Every kilogram of dry planting material can produce the fresh mushroom of 879.9g, and biological efficiency is up to 88%.
The foregoing is only preferred embodiment of the present invention, all equalizations done according to the present patent application the scope of the claims change and modify, and all should belong to covering scope of the present invention.
Claims (7)
1. an assay method for spot beautiful gill fungus bacterium bag degree of ripeness, is characterized in that: by the process of spot beautiful gill fungus bacterium sample packet, mensuration bacterium comprises the enzyme activity in the water yield, total sugar content, content of reducing sugar, soluble protein content and bacterium bag, measures spot beautiful gill fungus bacterium bag degree of ripeness.
2. the assay method of a kind of spot according to claim 1 beautiful gill fungus bacterium bag degree of ripeness, it is characterized in that: described sample handling procedure is: spot beautiful gill fungus bacterium bag is cultivated and started in 140 days for 60 days, got every 10 days weight difference within 2g and in 10 days weight reduce bacterium bag at the bacterium bag of 7g-9g as laboratory sample; Bacterium bag is divided into upper, middle and lower three part every 6cm, respectively compost is stirred, accurately take 10 g composts in 50 mL beakers, add 40 mL distilled water and shake up 2 h in 25 DEG C of shaking tables, get liquid in 50 mL centrifuge tubes, centrifugal 15 min of 9000 r/min, supernatant is crude enzyme liquid; Separately take 20 g in plate, dry to constant weight for 80 DEG C, measure water cut.
3. the assay method of a kind of spot according to claim 1 beautiful gill fungus bacterium bag degree of ripeness, is characterized in that: described
total sugar content assay method: accurately take the planting material of 0.25 g oven dry to constant weight in 10 mL centrifuge tubes, add 5 mL distilled waters and 1.5 mL concentrated hydrochloric acids, 3 h are hydrolyzed in boiling water bath, supernatant is poured into 50 mL conical flasks after leaving standstill cooling, rejoin 5 mL distilled waters and 1.5 mL concentrated hydrochloric acids, be hydrolyzed 2 h in boiling water bath, after leaving standstill cooling, supernatant be incorporated in 50 mL conical flasks, be settled to 50 mL with distilled water washing;
Get 1 mL after mixing and be again settled to 50 mL; Then centrifugal 15 min of 9000 r/min; Get 1 mL supernatant, add 1 mL 5 % phenol, add the 5 mL concentrated sulphuric acids fast, leave standstill 10 min, mixing, 30 water-bath 20 mi; 490 nm measure absorbance; Sample absorbance substitutes into the total sugar concentration of typical curve regression equation and derived sample; Total sugar concentration × Sample Dilution the multiple of sample total sugar concentration μ g/mL=Sample Dilution.
4. the assay method of a kind of spot according to claim 1 beautiful gill fungus bacterium bag degree of ripeness, is characterized in that: described
moisture determination method: after using calibration, pH meter measures supernatant and measures pH, and record plate original weight, weighs plate every day and sample general assembly (TW) no longer reduces to weight, illustrates and dries completely, and record data also calculate water cut.
5. the assay method of a kind of spot according to claim 1 beautiful gill fungus bacterium bag degree of ripeness, is characterized in that: described
content of reducing sugar assay method: accurately take planting material 0.5 g of oven dry to constant weight in 10 mL centrifuge tubes, add 5 mL distilled waters in boiling water bath, boil 2 h, then centrifugal 15 min of 9000 r/min, get 1 mL supernatant, add 1.5 mL DNS reagent, boiling water bath 7 min again, then use cold flow water cooling cessation reaction, be settled to 10 mL with distilled water, fully mix, under 540 nm, survey absorbance, and substitute into glucose standard curve regression equation and find corresponding glucose content.
6. the assay method of a kind of spot according to claim 1 beautiful gill fungus bacterium bag degree of ripeness, is characterized in that: described
soluble protein content assay method: get 1 mL supernatant, add the Coomassie brilliant G-250 working fluid of 5 mL, develop the color after 10 min, ultraviolet spectrophotometer measures 595 nm place light absorption values, distilled water is as blank, sample absorbance substitutes into the protein concentration that bovine serum albumin(BSA) typical curve regression equation tries to achieve sample, the protein concentration × Sample Dilution multiple of sample protein matter concentration μ g/ mL=Sample Dilution.
7. the assay method of a kind of spot according to claim 1 beautiful gill fungus bacterium bag degree of ripeness, is characterized in that: described
enzyme activity determination method in bacterium bag: under 520 nm, survey absorbance, and find corresponding glucose content from glucose standard curve, be calculated to be enzyme activity; Enzyme activity unit is defined as: the hydrolysis substrate per minute enzyme amount generated needed for 1 μ g glucose is defined as 1 enzyme activity unit " U " under these conditions.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410583701.2A CN104316478A (en) | 2014-10-28 | 2014-10-28 | Determination method of hypsizigus marmoreus fungi pack ripeness |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410583701.2A CN104316478A (en) | 2014-10-28 | 2014-10-28 | Determination method of hypsizigus marmoreus fungi pack ripeness |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104316478A true CN104316478A (en) | 2015-01-28 |
Family
ID=52371739
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410583701.2A Pending CN104316478A (en) | 2014-10-28 | 2014-10-28 | Determination method of hypsizigus marmoreus fungi pack ripeness |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104316478A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105717073A (en) * | 2015-09-30 | 2016-06-29 | 广东工业大学 | Cotton fiber maturity measuring method based on densitometry |
| CN109269934A (en) * | 2018-08-22 | 2019-01-25 | 漳州傲农牧业科技有限公司 | A kind of measuring method of Swine plasma protein water-soluble substance |
| CN109655414A (en) * | 2018-11-27 | 2019-04-19 | Oppo广东移动通信有限公司 | Electronic equipment, information-pushing method and Related product |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1397157A (en) * | 2002-08-09 | 2003-02-19 | 深圳市天蕈生物食品开发有限公司 | Culture medium and industrial production technology for Benzhandi mushroom |
| KR20060109142A (en) * | 2005-04-15 | 2006-10-19 | 최경환 | Variable hgpsizygus marmoreus and the method of cultivation |
| CN101589678A (en) * | 2009-07-01 | 2009-12-02 | 上海浦东天厨菇业有限公司 | The hypsizygus marmoreus produced by applying liquid strains original hase elicits method |
-
2014
- 2014-10-28 CN CN201410583701.2A patent/CN104316478A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1397157A (en) * | 2002-08-09 | 2003-02-19 | 深圳市天蕈生物食品开发有限公司 | Culture medium and industrial production technology for Benzhandi mushroom |
| KR20060109142A (en) * | 2005-04-15 | 2006-10-19 | 최경환 | Variable hgpsizygus marmoreus and the method of cultivation |
| CN101589678A (en) * | 2009-07-01 | 2009-12-02 | 上海浦东天厨菇业有限公司 | The hypsizygus marmoreus produced by applying liquid strains original hase elicits method |
Non-Patent Citations (4)
| Title |
|---|
| 左琦等: "食用菌总糖含量测定方法的研究", 《食用菌学报》 * |
| 袁胜东: "猴头菌生长过程中基质内营养物质含量和胞外酶活研究", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
| 高君辉等: "真姬菇培养时间与栽培料失重、含水量和产量的关系", 《食用菌学报》 * |
| 黄年来: "栽培真姬菇的日本技术", 《中国食用菌》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105717073A (en) * | 2015-09-30 | 2016-06-29 | 广东工业大学 | Cotton fiber maturity measuring method based on densitometry |
| CN109269934A (en) * | 2018-08-22 | 2019-01-25 | 漳州傲农牧业科技有限公司 | A kind of measuring method of Swine plasma protein water-soluble substance |
| CN109655414A (en) * | 2018-11-27 | 2019-04-19 | Oppo广东移动通信有限公司 | Electronic equipment, information-pushing method and Related product |
| CN109655414B (en) * | 2018-11-27 | 2021-11-02 | Oppo广东移动通信有限公司 | Electronic equipment, information pushing method and related product |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Klein et al. | A system of miniaturized stirred bioreactors for parallel continuous cultivation of yeast with online measurement of dissolved oxygen and off‐gas | |
| CN104316478A (en) | Determination method of hypsizigus marmoreus fungi pack ripeness | |
| Schlatmann et al. | Large-scale production of secondary metabolites by plant cell cultures | |
| ATE287537T1 (en) | MEASUREMENT OF METABOLIC CHANGES | |
| CN102994387A (en) | High throughput screening method of aminopeptidase and high-yield strain thereof | |
| CN110301291A (en) | A kind of breeding method of hickory chick strain | |
| Córdova-López et al. | Biomass estimation of Aspergillus niger growing on real and model supports in solid state fermentation | |
| Mercado-Flores et al. | Proteinases and exopeptidases from the phytopathogenic fungus Ustilago maydis | |
| CN111492895A (en) | Poplar sawdust composite culture medium suitable for hypsizigus marmoreus planting and preparation method thereof | |
| Boyle et al. | Development and comparison of methods for measuring growth of filamentous fungi on wood | |
| CN108138111A (en) | Monitor the state deviation in bioreactor | |
| CN104789635A (en) | Method for evaluating activity of aspergillus niger mouldy bran spore | |
| Nopharatana et al. | Use of confocal microscopy to follow the development of penetrative hyphae during growth of Rhizopus oligosporus in an artificial solid‐state fermentation system | |
| CN101589677A (en) | The training quality detection method of hypsizygus marmoreus produced by applying liquid strains | |
| CN102405762B (en) | Hypsizigus marmoreus spawn quality detection method | |
| CN109459431A (en) | The detection method of amylase in a kind of oyster sauce | |
| CN104195219A (en) | New method for detecting D-allose based on double-enzyme coupling high throughput | |
| CN110423794A (en) | A kind of method of Xianggu mushroom strain fruiting ability quick predict | |
| Dubey et al. | An enzyme-linked immunosorbent assay for the estimation of fungal biomass during solid-state fermentation | |
| CN102080120A (en) | Method for detecting quality change of hypsizigus marmoreus strains | |
| CN108077023A (en) | A kind of natural plants soilless culture nutrient fluid based on blue or green stalk leaching liquor | |
| CN216363436U (en) | Quantitative food fermentation and dough kneading fermentation device | |
| Batta et al. | Sucrose accumulation and expression of enzyme activities in early and mid-late maturing sugarcane genotypes | |
| CN109258306A (en) | A kind of elegant precious mushroom strain mating system rapidly and efficiently | |
| CN103636918B (en) | One is rich in linoleic modification protein isolate preparation method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20150128 |
|
| RJ01 | Rejection of invention patent application after publication |