50 results about "Aspergillus sojae" patented technology

The present invention relates to a mutant strain of Aspergillus sojae with an enhanced protease activity and a preparation method of a natural taste enhancer using the same. More specifically, the present invention relates to the mutant strain of Aspergillus sojae with an enhanced protease activity is obtained by selecting a strain with high protease activity and treating it with N-methyl-N′-Nitroso-N-nitrosoguanidine (NTG) and inducing a mutation through irradiating, and a preparation method of a natural taste enhancer using protein hydrolysate obtained by hydrolyzing the protein sources with their cultures.

The invention relates to a method for effectively removing residual aluminum in water by adopting a flocculation process, which adopts the flocculation process by using aluminum series flocculating agents as main flocculating agents and microorganism flocculating agents as auxiliary agents and comprises the following steps of: 1) selecting aluminum series flocculating agent polyaluminium chloride (PAC) as the main flocculating agents; 2) selecting microorganism flocculating agent soy aspergillus sojae strains as the auxiliary agents; and 3) adding the PAC and the microorganism flocculating agents into water to be treated, respectively carrying out fast stirring and slow stirring, and discharging supernatant fluid after stillness. The method has the advantages that the chemical flocculating agent and microorganism flocculating agent compounding treatment method is adopted, the problem of secondary pollution caused by the application of the PAC in the prior art is solved, the consumption of the PAC is reduced, the flocculation efficiency is improved, the effect of effectively removing the residual aluminum is reached through the bonding of the microorganism flocculating agents and the PAC and the immeshing effect of the microorganism flocculating agents, and the method belongs to an economic and efficient flocculation method of removing the residual aluminum in the water.

The invention discloses aromatic greengage filling, a production method thereof, and a greengage filling food. The method comprises the following steps: greengage is cleaned, denucleated and subjected to pulping treatment, biological enzyme treatment by flavoring enzymes, finish grinding and superhigh pressure homogenization treatment; the conditions of the biological enzyme treatment are that the temperature is between 30 and 60 DEG C, PH value is 4 to 6, and the time is 1 to 6 hours; the flavoring enzymes take two to three of Aspergillus niger, Aspergillus oryzae, Aspergillus ficcum, Aspergillus fumigatus, Aspergillus japonicus Saito, Aspergillus awamori, Aspergillus carbonarius, Aspergillus terreus, Aspergillus tubingensis, Aspergillus usammi and Aspergillus sojae as zymogens; and the greengage is subjected to mixed fermentation in a culture medium for 2 to 5 days at a temperature of between 20 and 40 DEG C and under the condition of 200 to 600r / min, and leached, filtrate is leached and centrifuged, and the supernatant is taken as flavoring enzyme liquid. By the production method, high-quality aromatic greengage filling food can be manufactured, and has unique flavor, aromatic and rich fragrance of the greengage, high quality and wide development prospect.

The invention relates to a preparing method of a corn gluten polypeptide for fodder, belongs to the technical field of processing of fodder and particularly relates to a method of preparing the corn gluten polypeptide for the fodder by taking corn gluten meal as a raw material. The method comprises the following steps: swelling the corn gluten meal, then adding water, uniformly stirring, adjusting pH to 7.0-7.2, sterilizing, cooling to 45-50 DEG C, adding a neutral protease, performing hydrolysis for 6-8h, boiling, cooling to 26-28 DEG C, inoculating aspergillus sojae, fermenting for 26-30h, centrifuging to obtain clear liquid, concentrating the clear liquid and spray-drying to obtain a finished product. The preparing method of the corn gluten polypeptide for the fodder, disclosed by the invention, effectively integrates the respective advantages of a swelling method, an enzyme method and a fermentation method; the corn gluten polypeptide for the fodder is rich in nutrients and has prominent effects; furthermore, the prepared gluten polypeptide has no bitter taste, no peculiar smell and good palatability.

The invention discloses a compound microbial agent for anaerobic digestion treatment of sludge and a production method of the compound microbial agent. The compound microbial agent is a liquid microbial agent which is prepared through fermentation culture and comprises the following bacteria: bacillus subtilis, streptomyces albus, streptomyces griseus, candida utilis, aspergillus sojae, lactobacillus bulgaricus, lactococcus lactis, rhodopseudomonas palustris, rhodopseudomonas capsulate and rhodobacter sphaeroides, and the number of total viable bacteria in 1ml of the compound microbial agent is not less than 2*10<8>. All the bacteria used by the compound microbial agent have metabiosis and symbiosis effects and form a biodegradation system with very high enzymatic activities. When the compound microbial agent disclosed by the invention is used for carrying out anaerobic digestion treatment on sludge, the degradation rate of organic matters can be effectively increased, the sludge hydrolytic process can be accelerated, the anaerobic digestion time of the sludge can be shortened, and the biological gas yield can be increased.

The invention discloses a compound microorganism combination microbial agent, a preparation method and application thereof, a sludge treatment method for a black-odor river. The compound microorganism combination microbial agent comprises the three kinds of microbial agent combinations of a microbial agent A combination, a microbial agent beta combination and a microbial agent gamma combination, and the microbial agent A combination comprises the microbial strains of bacillus coagulans, lactic acid bacteria, saccharomycetes, aspergillus sojae and thiobacillus denitrificans; the microbial agent beta combination comprises the microbial strains of lactic acid bacteria, bacillus subtilis, photosynthetic bacteria and saccharomycetes; and the microbial agent gamma combination comprises the microbial strains of lactic acid bacteria, bacillus subtilis, saccharomycetes, bacillus licheniformis and bacillus coagulans. The compound microorganism combination microbial agent can be used for treatment of bottom mud of the black-odor river, the treatment period is short, the thickness of the bottom mud can be effectively reduced, the content of harmful matters in the bottom mud is reduced, the situation that nitrogen and phosphorus of a black and odorous water body exceed a standard can be solved, and odorous gas hydrogen sulfide in the water body can be eliminated.

The invention discloses a pretreatment method for enhancing anaerobic sludge digestion by virtue of a compound microorganism bacterium agent. The pretreatment method comprises the following steps: concentrating residual sludge, adding a compound microorganism bacterium agent, performing stirring reaction, inoculating the pretreated residual sludge into an anaerobic digestion pond according to a certain ratio, and performing mesophilic anaerobic digestion on anaerobic sludge, wherein the compound microorganism bacterium agent is prepared by mixed fermentation of 5%-20% of each of bacillus subtilis, streptomyces albus, streptomyces griseus, candida utilis, aspergillus sojae, lactobacillus bulgaricus, lactococcus lactis, rhodopseudomonas palustris, rhodopseudomonas capsulate and rhodopseudomonas spheroides, the total viable count is not less than 2*108 / ml, and the pH value of the compound microorganism bacterium agent is 3-4. According to the method, the sludge hydrolysis process can be accelerated, the sludge retention time can be shortened, the biological gas production rate can be increased, and the anaerobic digestion efficiency can be improved.

The present invention is to provide a method for producing a material which contains xylo-oligosaccharides being rich in xylobiose and xylotriose, an antioxidant active substance and a dietary fiber, and which is useful as a food material and a health supplement, by a simple process, with efficiency, and at a low cost. The method comprises the steps of: subjecting a hemicellulose-containing plant resource such as bagasse to a steam treatment followed by an explosion treatment (2.5 Mpa, 70 seconds); mixing wheat bran into this steamed and exploded material; adjusting water content of this mixture to about 50%; subsequently, preparing koji with the use of a koji fungus having a high xylanase activity obtained by conducting single spore isolation from Aspergillus sojae (NFRI1147); adding water to the koji thus obtained; mixing the steamed and exploded material into a koji dispersion wherein koji is dispersed in water; adjusting water content of this mixture to about 50%; and subsequently, fermenting the mixture. After conducting the fermentation, a treatment to terminate the fermentation is conducted and the resultant is dried and then finely ground. As the steamed and exploded material to be mixed into the koji dispersion, 8- to 10-fold amount of the steamed and exploded material used for the koji preparation, is used.

The invention discloses a method for increasing utilization rate of soybean sauce raw materials ultrasonically and catalyzing maturation of flavor, and belongs to the technical field of processing ofcondiments. The method comprises the following steps: preparing soy leaven by taking aspergillus oryzae or soybean sauce aspergillus sojae as fermentation culture and taking bran as a raw material, wetting, cooking and cooling soybeans / soybean meal / wheat / bran and then inoculating bran soy leaven to prepare yeast for making hard liquor; and after the yeast for making hard liquor falls into a tank or a pool or a cylinder and is mixed with saline completely, performing fermentation, performing at least one turn of ultrasonic treatment at the early stage and the later stage of fermentation to fulfill the aims of increasing the utilization rate of the soybean sauce raw materials and catalyzing the maturation of the flavor. Compared with the control soybean sauce produced under the same condition, the soybean sauce produced by the method of the invention has typical soybean sauce flavor and color; the raw material utilization rate and the amino acid state content of the soybean sauce produced by the method are increased by 5 to 10 percent and 5 to 7 percent correspondingly compared with the those of the control; and the fermentation cycle of high-salt loose-state soybean sauce is shortened by more than 60 days, the flavor and the fragrance of the high-salt loose-state soybean sauce and the low-salt solid-state soybean sauce are obviously improved, and the precipitate forming quantityis reduced.

The present invention in particular relates to a preparation method of biological type sludge conditioning agent and the application thereof. The biological type sludge conditioning agent is prepared by using aspergillus sojae (Aspergillus sojae) as raw material, and the obtained sludge conditioning agent can be applied to the sludge dewatering. The coagulation activity of the conditioning agent of the present invention is high, the cost is low, the dosage output is high, the stability is strong, the applicable range is wide, and the present invention has the biological degradability and the environmental friendliness, and the second pollution is not caused.

The invention discloses a compound microorganism combination microbial agent, a preparation method and application thereof, a comprehensive treatment method for a black-odor river. The compound microorganism combination microbial agent comprises the three kinds of microbial agent combinations of a microbial agent A combination, a microbial agent C combination and a microbial agent gamma combination, and the microbial agent A combination comprises the microbial strains of bacillus coagulans, lactic acid bacteria, saccharomycetes, aspergillus sojae and thiobacillus denitrificans; the microbial agent C combination comprises the microbial strains of bacillus coagulans, rhizopus oryzae, saccharomycetes, aspergillus sojae and penicillium chrysogenum; and the microbial agent gamma combination comprises the microbial strains of lactic acid bacteria, bacillus subtilis, saccharomycetes, bacillus licheniformis and bacillus coagulans. The compound microorganism combination microbial agent can be used for comprehensive treatment of the black-odor river, the treatment period is short, the situation that nitrogen and phosphorus of a black and odorous water body exceed a standard can be solved, and odorous gas hydrogen sulfide in the water body can be eliminated.

The invention discloses a microbial sewage treatment agent. The microbial sewage treatment agent is prepared from the following raw materials by weight: 15-25 parts of zeolite powder, 25-35 parts of activated carbon, 5-15 parts of potassium feldspar, 5-15 parts of lauryl trimethyl ammonium bromide, 5-15 parts of magnesium sulfate, 1-5 parts of Rhodococcus erythropolis, 1-5 parts of Phanerochaete chrysosporium, 1-5 parts of Aspergillus sojae, 1-5 parts of Thiobacillus and 5-15 parts of potassium phosphate. The microbial sewage treatment agent of the invention is prepared from environmentally friendly raw materials, and is high in the specific surface area of an inorganic filler, high in activity, high in dispersibility and strong in adsorptivity; at the same time, the microbial sewage treatment agent can play the roles of demulsification, flocculation, bridging and foaming; and when applied to treatment of sewage from textile mills, the microbial sewage treatment agent has obvious treatment effect and good stability and is hardly affected by the environment.

The invention relates to a processing method of foodstuff, in particular to a production method of low-salt maize soy sauce by fermentation. The first step is steaming materials, namely, the maize is steamed and cooked; the second step is making starters, namely, mold starters with the amount accounting for about 0.3 percent of that of materials are added into the maize at the temperature of 35-45 DEG C; the third step is fermentation; the fourth step is leaching and the fifth step is preparation and processing: raw soy sauce is heated and sterilized for conventional preparation and is then filtered after standing for more than three days; the soy sauce is then packaged to obtain finished products. By adopting the maize as the material, aspergillus (aspergillus oryzae or aspergillus sojae) with pure culture for making starters and low-salt solid state fermentation, the invention can improve the flavor and the quality of the soy sauce; the sauce flavor is enhanced by adding dilute syrup liquid which is conducive to microorganisms of yeast, etc., when in fermentation. The invention also adopts modern fermentation techniques and fully-sealed and mechanized production, thereby having sanitary and reliable products, high labor productivity and materials utilization rate, relatively low cost as well as high-level nutrition, taste and quality.

To promote the digestion of an animal to thereby elevate the feed efficiency or prevent or ameliorate intestinal infections such as inflammatory intestinal injury of the animal, a combination of at least one fungus selected from Aspergillus sojae, Aspergillus tamarii, Aspergillus foetidus, Aspergillus niger, and Aspergillus oryzae with an acidic enzyme produced by the fungus is administered to the animal.

The present invention discloses thick broad-bean sauce fermented by mixed pure strains and a preparation method thereof. The method consists of the following steps: firstly, water is added to soak shelled dry broad bean halves to enable the broad bean halves to be fully swollen by absorbing water until no white hard hearts appear on the fracture surfaces; then the soaked wet broad beans are drained, the drained broad beans are put into a steaming pot to be steamed until the surface layer shows gas, a lid covers the steaming pot, then fire is turned off, the broad bean halves are stewed, and the stewed broad bean halves are taken out from the pot; after the steaming of the materials, the steamed broad bean halves are cooled to a room temperature at a sterile condition, aspergillus oryzae spore powder, aspergillus sojae spore powder and flour are inoculated into the broad bean halves, and the materials are mixed well; the inoculated broad bean halves are cultured in an incubator to prepare koji; and finally the koji and sterile saline is added to conduct fermentation, and the mixture is fermented in the incubator to prepare the thick broad-bean sauce. The pure strain fermentation avoids the contamination of hybrid strains, improves the success rate of fermentation and enhances consumption safety. Besides, the fermentation only needs 20-30 d, the fermentation time is shortened, and the product is stable in quality. All the advantages indicate that the preparation method is suitable for mass production.

The present invention discloses a tea leaf caramel color preparation method which comprises the following steps: 1) firstly picked fresh tea leaves are rinsed, cooked at a high temperature of 100-120 DEG for 3-5 minutes, cooled to room temperature, and subjected to a sealed fermentation; 2) flour is added with aspergillus sojae, the mixture is stirred evenly, the firstly fermented tea leaves obtained in the step 1 is fully stirred evenly with the flour, the mixture is fermented in the shade until the mixture is completely black and has a soy sauce fragrant taste, and the fermentation is completed. The advantages of the caramel colored tea leaves are that the tea leaves have pure and natural caramel colors, and are rich in nutrients.

Disclosed is a safe and convenient means for supporting the digestive activity in an animal and increasing the feed efficiency. Specifically, disclosed is a means for preventing / treating an infectiousdisease by inhibiting the proliferation of a pathogenic bacterium or a coccidium in the intestine of the animal and, consequently, increasing the body weight of the animal. At least oneAspergillus bacterium selected from Aspergillus sojae, Aspergillus tamarii, Aspergillus foetidus,Aspergillus niger and Aspergillus oryzae, a culture containing an acidic enzyme produced by the bacterium, and Bacillus subtilis are administered to an animal in specified amounts.

The invention refers to a method for manufacturing peptide product for promoting calcium absorption, comprising the steps of: mixing raw materials including soy protein with water to form a mixture including 45%-65% weight percentage water content; inoculating Aspergillus oryzae, Aspergillus sojae or their aspergillus flora into the mixture to obtain a culture, wherein an inoculum size of the aspergillus flora accounts for 0.006%-0.05% of soy protein raw materials; perform solid state fermentation on the mixture with a relative humidity of 85%-95% and a fermentation temperature between 23 and 35 DEG C to obtain a fermentation product; performing hydrolysis reaction on the fermentation product to obtain soybean hydrolysate; clarifying processing the soybean hydrolysate and collecting hydrolysate to obtain peptide products for promoting calcium absorption.

The purpose of the present invention is to provide a strain having a significantly increased frequency of homologous recombination, which is necessary for gene disruption of gene replacement by gene targeting of a mitosporic filamentous fungus. This invention relates to a transformant having an increased frequency of homologous recombination (gene targeting), which is a mitosporic filamentous fungus belonging to Trichocomaceae, due to suppression of a ku gene by ku gene disruption or antisense RNA method, to a method for the production of a gene-disruption stain, gene-deletion strain, gene-replacement strain, gene-insertion strain or chromosome-modification strain by means of the gene targeting method using said transformant, and to the above-mentioned ku genes such as those derived from Aspergillus sojae and Aspergillus oryzae, and their expressed products (proteins).

A method for preparing soybean peptide-calcium chelate is disclosed. The method of the invention includes mixing a material containing soy protein with water to form a mixture having a water content of about 45-65% by weight, and inoculating the mixure with Aspergillus oryzae, Aspergillus sojae, or the combination thereof in an amount of about 0.006% -0.05% based on the total weight of the soy protein material. Then, the resulting culture is subjected to a solid state fermentation to obtain a fermentation product. After the fermentation product is hydrolyzed, the hydrolysate is subjected to clarification, and then calcium ion is mixed with the clarified hydrolysate with a molar ratio of 2: 1 to 0.5: 1, followed by adjusting the water content and pH to obtain a soybean peptide-calcium chelate solution.

PURPOSE: A method for fermenting wild ginseng and fermented wild ginseng manufactured by the same are provided to maximize a physiologically active function of the wild ginseng by increasing ginsenoside. CONSTITUTION: Wild ginsengs are cleaned and seed germs are sprayed and cultivated. The cultivated ginsengs are packed in a vacuum state. The vacuum packed wild ginsengs are fermented. The fermented wild ginseng is heated and sterilized at 60 to 65 degrees centigrade for 30 to 60 minutes and is cooled at 4 to 10 degrees centigrade as an intermittent sterilization. The seed germ is made of Aspergillus oryzae, Aspergillus sojae, Aspergillus shirousamii, Rhizopus sp., or Zygomycetes. The cultivation is performed at a temperature of 35 to 45 degrees centigrade and a humidity condition of 60 to 85 %RH for 1 to 4 hours.

The invention discloses a method for fixing flocculated and settled lipid yeast through microbial flocculant cooperating with activated carbon, belongs to the technical field of microorganisms, aims at utilizing characteristics of the activated carbon combined with microbial flocculation technology to flocculate and settle the lipid yeast and is is a method for fixing the flocculated and settled lipid yeast through the microbial flocculant cooperating with diatomite. The method includes the steps of treatment of the activated carbon, preparation of the microbial flocculant through aspergillus sojae, preparation of a Starkeyi lipid yeast culture solution, immobilization of Starkeyi lipid yeast through the activated carbon, and flocculation and settling of the Starkeyi lipid yeast immobilized by the activated carbon through the microbial flocculant. The immobilized Starkeyi lipid yeast is simple in operation and high in flocculation and settling rate which is maintained between 93.90% and 99.62%.

To promote the digestion of an animal to thereby elevate the feed efficiency or prevent or ameliorate intestinal infections such as inflammatory intestinal injury of the animal, a combination of at least one fungus selected from Aspergillus sojae, Aspergillus tamarii, Aspergillus foetidus, Aspergillus niger, and Aspergillus oryzae with an acidic enzyme produced by the fungus is administered to the animal.

The invention discloses a production method of nutritional selenium-enriched soy sauce and belongs to the technical field of fermentation engineering. The production method includes three steps: making koji, fermenting and processing to obtain the soy sauce. Total selenium content of the soy sauce is 2.2-2.4mg / L, and organic selenium content is 1.6-1.8mg / L, so that the soy sauce is rich in selenium content. Total acid of the soy sauce prepared through synergism of aspergillus oryzae, aspergillus sojae and saccharomyces soya is 1.70-1.72g / 0.1L, reducing sugar content is 6.38-6.41g / 0.1L, total nitrogen content is 1.83-1.86g / 0.1L, and the soy sauce is good in fresh taste, coordinated in flavor and good in fragrance and has better fermentation quality than that when one of aspergillus oryzae or aspergillus oryzae and aspergillus sojae or saccharomyces soya is used alone.

The invention discloses a soy sauce brewing method for increasing an amino acid yield by utilizing an enzyme preparation. The soy sauce brewing method comprises the following steps: mixing aspergillus oryzae and / or aspergillus sojae with cooked soybean and starchy auxiliary materials, and then fermenting to prepare soy sauce koji; mixing the obtained soy sauce koji with salt water to prepare soy sauce mash, and then fermenting in nature to obtain soy sauce; adding the enzyme preparation which accounts for 3 percent to 10 percent of the total weight of a koji making material into the soy sauce mash, wherein the enzyme preparation is obtained by fermenting aspergillus niger and desalted sauce residue, and acid proteinase activity of the enzyme preparation is 2000 U / g or above. According to the method disclosed by the invention, by adding the enzyme preparation during fermentation, an enzyme system lacking in the aspergillus oryzae and / or aspergillus sojae is remedied, the acid proteinase activity of the soy sauce mash is increased, amino acid content in natural soy sauce is increased, occurrence of Maillard reaction is promoted, and a soy sauce flavor is promoted.

The invention provides a production method of sweet seasoning oil. The production method comprises the following steps of (1) mixing and stirring flour with water, performing kneading to obtain dough,performing dicing, and performing steaming to obtain cooked dough dices for standby application; (2) pouring and sprinkling aspergillus sojae to the cooked dough dices obtained through steaming, performing reversing and stirring, and beginning to mildew to obtain sauce blanks, wherein the mildewing environment temperature is 28-35 DEG C, and the humidity is smaller than 10%; (3) compounding saltwater with pure water and refined salt, wherein the concentration of the salt water is 15-18%; (4) firstly, putting a soy sauce pump at a central position of a soy sauce jar, and then putting the sauce blanks around the soy sauce pump; (5) on the same day of putting the blanks, pouring and sprinkling appropriate salt water to the soy sauce pump, and at the later stage, pouring and sprinkling the salt water in a manner of pouring and sprinkling the salt water once every 4 days, pouring and sprinkling the salt water once every 6 days, pouring and sprinkling the salt water once every 8 days and pouring and sprinkling the salt water once every 16 days until oil appears; and (6) introducing the oil obtained in the step (4) into an indoor oil cylinder for sealed storage for 2-3 months, and performing secondary fermentation so as to obtain finished products. The sweet seasoning oil made by the method for making the sweet seasoning oil provided by the invention is fresh and delicious in flavorand red in color.

The invention relates to the technical field of sewage treating, in particular to a sewage treating agent. The sewage treating agent is prepared from 20-35 parts of a microorganism treating agent and 10-20 parts of limestone. White rot fungi and serpula lacrymans schroet are mixed according to the weight ratio of (1-3):(1-3), aspergillus sojae and rhodococcus erythropolis are mixed according to the weight ratio of (1-3):(1-3), the two mixtures are mixed according to the weight ratio of 1:1 to obtain the microorganism treating agent, and the microorganism treating agent is mixed with the limestone to prepare the sewage treating agent. The sewage treating agent is used for treating textile mill sewage and is obvious in treating effect, good in stability and little in influence generated by environment.

The invention discloses a pretreatment method for enhancing anaerobic sludge digestion by virtue of a compound microorganism bacterium agent. The pretreatment method comprises the following steps: concentrating residual sludge, adding a compound microorganism bacterium agent, performing stirring reaction, inoculating the pretreated residual sludge into an anaerobic digestion pond according to a certain ratio, and performing mesophilic anaerobic digestion on anaerobic sludge, wherein the compound microorganism bacterium agent is prepared by mixed fermentation of 5%-20% of each of bacillus subtilis, streptomyces albus, streptomyces griseus, candida utilis, aspergillus sojae, lactobacillus bulgaricus, lactococcus lactis, rhodopseudomonas palustris, rhodopseudomonas capsulate and rhodopseudomonas spheroides, the total viable count is not less than 2*108 / ml, and the pH value of the compound microorganism bacterium agent is 3-4. According to the method, the sludge hydrolysis process can be accelerated, the sludge retention time can be shortened, the biological gas production rate can be increased, and the anaerobic digestion efficiency can be improved.

The present invention relates to an Aspergillus non-inherited genetic variant having enhanced protease activity, and to a production method for a natural flavour enhancer employing the same, and more specifically relates to: an Aspergillus non-inherited genetic variant having enhanced protease activity which is obtained by selecting a strain having a high level of protease activity inducing genetic variation by treating the same with NTG (N-methyl-N'-nitroso-N-nitrosoguanidine) and exposing the same to radiation; and to a production method for a natural flavour enhancer employing a hydrolysed protein obtained by culturing the Aspergillus non-inherited genetic variant, adding the resulting culture product to a protein source and allowing hydrolysis to proceed.