CN1156178A - Biological deodorization method for silkworm pupa after reeling silk - Google Patents

Abstract

A biological deodorization method for silkworm pupa after reeling silk includes mixing the pupa mash or dry pupa powder with wheat bran, rice husk and water, steam sterilizing at 110-121 deg.C, inoculation of aspergillus As3.374 culture medium, fermentation at 33-35 deg.C and pH of 7.0-7.2 for 58 hr. or more, and keeping temp. constant at 30 deg.C for 2 hr. Processed silk pupa has protease activity of over 4000 units and trimethylamine content less than 0.15%.

Description

The biological deodorization method of silk reeling pupa

The present invention relates to the deodorization technique of silk reeling pupa, particularly remove the method for protein of silk filature pupa matter stink with biotechnology.

Silk reeling pupa is large renewable resource of silkworm industry, when silk cocoon filature system one is given birth suddenly silk, can obtain the dried pupa about a ton.The pupa powder contains 48.9% pupa albumen; 29.5% fat and other abundant nutrition; aspect food, medicine, the light textile extensive use is being arranged; be to produce peptone, protein powder, moriamin-s; foodstuff additive amino acid peptide, natural health care, feed, pupal fat, daily use chemicals base-material acylated peptide, leather smoothing agent, the raw material of light textile solubility promoter and washing composition, still; because silk reeling pupa deodorization technique difficulty directly affects pupa resource comprehensive processing series product quality.The urgent need of producing under the pressure of silk silk fabric industry, the protein of silk filature pupa deodorization technique improves and still rests on general chemical technique and the equipment, breakthrough is not seen in the deodorization of Applied Biotechnology fermentation silk reeling pupa, therefore demands carrying out the research of removing the silk reeling pupa stink with biotechnology urgently.

Protein of silk filature pupa is smelly, mainly be because a large amount of amine substances are arranged, as Trimethylamine 99, diethylamine, normal hexyl Amine, particularly Trimethylamine 99, just can deodorization so remove or reduce amine content.

The biological deodorization method that the purpose of this invention is to provide a kind of silk reeling pupa, the deodorization of biological fermentation silk reeling pupa makes protein of silk filature pupa front three amine content＜0.5%.

Realize that the above-mentioned purpose technical scheme is that pupa gruel of silk reeling pupa water or dry pupa powder are broken into powder, cross 20 mesh sieves, add wheat bran, cavings and water are made substratum after the sterilization, the multiplication culture thing of inoculation As3.374 aspergillus terricola, heating fermentation, filter deodorization solubility pupa albumen, filter residue returns fermentation again.The concrete grammar step is:

1, by silk reeling pupa: wheat bran: cavings=100: (8.75-20.0): (1.25-2.00) siccative part by weight weighing batching, tap water, the sealing of adding dosage 100%-120%,

2,110-121 ℃ of steam sterilizing 30 minutes, make substratum.

Inoculation when 3, temperature to be expected drops to 30-31 ℃, As3.374 culture solid seed, add-on is 4-8% weight (is radix with the substratum).

4, at 33-35 ℃, PH7.0-7.2 fermentation 〉=58 hours.

5,30 ℃ of constant temperature 2 hours, promptly finish deodorization.

Sampling records proteinase activity more than 4000 units, residual amine amount＜0.15%.

The dried pupa of its raw material filature, the pupa gruel of filature water and wheat bran are provided by Luotian, Hubei Province star's industry main office, and cavings is bought on the market.As3.374 multiplication culture thing is the As3.374 aspergillus terricola that Chinese canonical biometric preservation center provides, and the original seed bacterium carries out the product of multiplication culture on the inclined-plane seed culture medium.Slant medium: 25 gram dry pupa powders add 100 ml waters, boil 15 minutes, filter to get filtrate 80 milliliters, add the NaNO of this liquid measure ₃0.2%, K ₂HPO ₄0.1%, KCl0.01%, MgSO ₄0.05%, FeSO ₄0.001%, sucrose 3%, and agar 1.5-2% adds water to 100 milliliter.Insert the former bacterium of As3.374, cultivated 3-4 days down, obtain outward appearance and do not have the crape folding, bacterium colony is big at 30-32 ℃, the drabon look of thallospore, the microscopy spore is many, the thick shape of mycelia, neatly.

Monitoring method:

Proteinase activity: Fehling method

Acidity: 0.1N NaOH solution titration

PH value: measure with accurate PH (9-12) test paper

Residual amine: liquid phase chromatography is Lc post-ISC-07/S1504 with Tianjin, island LC-3A liquid chromatograph, look post, the photometry of FLD-1 fluorescent detector, condition: current-carrying speed 0.5 ml/min, pressure 60-70 kilograms per centimeter ², 55 ℃ of column temperatures are got base-material 5 gram damping fluids (PHg), and 40 ℃, 500 milliliters leached 1 hour, and it is to be measured to get 50 milliliters of leach liquors, and every sweathouse is every sampling monitoring in 24 hours once.

Fermentation container: disc type bio-reactor.

Accompanying drawing has provided the variation that bacteria growing produces enzyme and Trimethylamine 99 in the fermenting process.

Fig. 1 is that bacteria growing produces enzyme and Trimethylamine 99 consumption curve.

Fig. 2 is acidity and a PH curve in the fermenting process.

Embodiment 1:

Take by weighing 53.3 filature dry pupa powder, the 36.6 gram brans, 10.1 that restrained 20 mesh sieves and restrain cavings, add 500 ml waters, mix 500 milliliters of triangular flasks, 150 grams of packing into thoroughly, the envelope bottle is with 1 kilograms per centimeter ²Steam sterilizing 30 minutes, in the sterilising chamber, get 1 spore grow luxuriant honey As3.37 culture bacterial classification, wash into spore suspension for 10 milliliters with sterilized water, aseptic straw is drawn 1 milliliter, insert in the triangular flask material and mix thoroughly, inserting 30-33 ℃ of incubator cultivated 12 hours, shake bottle, shake bottle after 6 hours once more, when treating the mycelial growth caking, the button bottle becomes piece, cultivates after 30 hours, when producing a large amount of brown spore, be warmed up to 34-35 ℃ and cultivated 24 hours, survey proteinase activity more than 5000 units, the front three amine is below 0.05%.

Embodiment 2:

Take by weighing 75.2 gram filature water pupa system gruels, add wheat bran 14.8 grams, add 5 gram cavings, 120 milliliters in water is mixed 500 milliliters of triangular flasks, 100 grams of packing into thoroughly, the envelope bottle, 110-121 ℃ of sterilization 30 minutes, insert 4 milliliters of As3.374 bacterium spore suspensions with aseptic straw, insert 30-33 ℃ of incubator and tiltedly put and shake bottle after 8 hours and stand up, shake bottle after 6 hours again, after being cultured to 2 days, the button bottle is warming up to 35 ℃ and cultivated one day, records more than proteinase activity 4000 units, and the front three amine is below 0.15%.

Embodiment 3:

Take by weighing protein of silk filature pupa powder 26.7 grams, smart wheat bran 62.8 grams add 700 milliliters in cavings 16 grams and water, mix 500 milliliters of triangular flasks, 250 grams of packing into thoroughly, envelope bottle, 1 kilograms per centimeter ²Steam sterilizing 30 minutes, insert 0.5 milliliter of As3.374 bacterium spore suspension, insert 28-30 ℃ of incubator and tiltedly put cultivation 6 hours, shaking bottle directly puts, is warming up to 32 ℃ and cultivate after 12 hours and shake bottle again, the button bottle heats up dry after being cultured to 2 days, survey its proteolytic enzyme in 4000-5000 unit, the front three amine is below 0.01%.

Embodiment 4:

In the disc type bio-reactor, feed intake 10 kilograms under the normal temperature, use 1 kilograms per centimeter by the dried pupa 85% of solid medium weight, wheat bran 10%, cavings 5%, adding 100%-120% water ratio ²Steam sterilizing when waiting to expect temperature drop to 30-31 ℃ then, connects the As3.374 culture, and inoculum size is that material is measured 4% weight, heaps earlier and cultivates 5-6 hour, turning during product temperature rise to 35 ℃, 1.5 kilograms on every dish (siccative).Bring up to 31-33 ℃ with temperature of reactor this moment, continue to cultivate 7-10 hour, draw dish, the silk reeling pupa culture material is divided into fritter, is provided with sufficient air, be beneficial to the spore growth, cultivate and after 50 hours temperature of reactor is risen to 34-35 ℃, make it progressively dry, to 72 hours, the spore growth was evenly dense.Its thalli growth produces enzyme and Trimethylamine 99 consumption such as Fig. 1,1 expression proteolytic enzyme change curve among the figure, 2 residual amine Trimethylamine 99 change curves, as seen from Figure 1, the zymophyte growth reached in 48 hours produces the enzyme peak, and the mycelium reproduction speed is the highest, makes Trimethylamine 99 stink thing composition be line style rapidly and descends.Produce enzyme peak later stage amine spending rate and reach maximum value, fermentation end of a period front three amine reaches 0.05%.Illustrate when mycelial growth is vigorous, consumed a large amount of amines, reached and removed the smelly purpose of pupa.

At fermentation variation such as Fig. 2 except that acidity in the smelly process of pupa and pH value, 3 is change curves of acidity among the figure, the 4th, the pH value change curve, as can be seen, the variation of pH value is little during the fermentation, and acidity improves with the growth of fermentation time, but increase rate is little, increase acid generally in 3.5, illustrate that sterilization does not influence silk reeling pupa fermentation deodorising process.

Proving from above result, is feasible modern biotechnology method with the As3.374 native inulinase fermentation silk reeling pupa deodorizing technology technology of dwelling, and its technology is simple, and cost is low, and negative value added height is good in economic efficiency.

Claims (4)

1, a kind of biological deodorization method of silk reeling pupa is characterized in that:

Press silk reeling pupa: wheat bran: cavings=100: (8.75-200): (1.25-2.00) siccative part by weight weighing batching, add the heavy 100%-120% water of batching siccative, sealing, at 110-121 ℃ of steam sterilizing, inoculation As3.374 culture when temperature to be expected drops to 30-31 ℃, add-on is for the 4-8% of material weight, at 33-35 ℃, PH7.0-7.2 fermentation 〉=58 hours is then 30 ℃ of constant temperature 2 hours.

2, deodour method according to claim 1 is characterized in that 110-121 ℃ of steam sterilizing 30 minutes.

3, deodour method as claimed in claim 1 is characterized in that the As3.374 culture is that aspergillus terricola As3.374 is on slant medium, at the 30-32 ℃ of culture of cultivating 3-4 days down.

4, deodorizing method as claimed in claim 3 is characterized in that described slant medium, is that 25 gram dry pupa powders add 100 ml waters, boils 15 minutes, filtration, adds the NaNO of this liquid weight in the filtrate ₃0.2%, K ₂HPO ₄0.1%, KCl0.01%, MgSO ₄0.05%, FeSO ₄0.001%, sucrose 3%, and agar 1.5-2% adds water to 100 milliliter.

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