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(2005). Changes in the colony morphology indicated the modification of few genes in the genome. 1b). (2014). In addition, M6 demonstrated the colony morphology of convex. Z., Sun, L. J., and Wang, Y. L. (2013). The colony morphology of many mutant spores on PDA plates differed from that of strain 3.042 due to the effect of ARTP treatment. (2019). Pendant la culture, l’Aspergillus oryzae, appelé « Tane-koji », crée un duvet blanc autour … Food Microbiol. Comparative genomic analysis of Aspergillus oryzae strains 3.042 and RIB40 for soy sauce fermentation. When the spores were treated for 240 s under the same conditions, nearly none of the spores survived. The acidic protease was purified from 4-day cultivation of Aspergillus oryzaeBCRC 30118 by DEAE Sephacel and Sephacryl S-200 HR chromatographs. Random mutagenesis of Clostridium butyricum strain and optimization of biosynthesis process for enhanced production of 1,3-propanediol. Une activité protéasique de 2635 U et une biomasse de 15,42 g sont obtenues au bout de 52 heures de culture. A study of chemical characteristics of soy sauce and mixed soy sauce: chemical characteristics of soy sauce. Aspergillus oryzae is known to have remarkable potential for the production of secreted enzymes. The specific activity, yield and purification fold were 117.62 kU/mg, 15.1% and 6.6, respectively. (2015a). 23, 397–402. (2010). doi: 10.1007/s00449-019-02079-9. The optimum temperature range for activity was 55–60°C, and the enzyme was stable at temperatures below 45°C. doi: 10.1007/978-3-662-46318-5_30, Xu, D., Pan, L., Zhao, H., Zhao, M., Sun, J., and Liu, D. (2011). The enzyme was purified by precipitation with tannic acid, chromatography on Duolite A-2, and gel filtration on Sephadex G-100. Figure 6. Biotehnol. J. Biosci. Antioxidant activities from the DPPH and ORAC assays and linearity ranges for the ORAC assay (net AUC versus concentration) performed on the control and hydrolysate samples produced by the protease from, Department of Food Science, School of Food Engineering, University of Campinas, 80 Rua Monteiro Lobato, 13083-862 Campinas, SP, Brazil, Optimum pH and temperature for protease activity, Optimum pH and temperature for protease stability, Optimum pH and temperature for the protease activity, pH and temperature for the protease stability, Kinetic parameters for irreversible thermal denaturation, T. A. S. Silva, A. Knob, C. R. Tremacoldi, M. R. Brochetto-Braga, and E. C. Carmona, “Purification and some properties of an extracellular acid protease from, V. Ramakrishna, S. Rajasekhar, and L. S. Reddy, “Identification and purification of metalloprotease from dry grass pea (, M. B. Rao, A. M. Tanksale, M. S. Ghatge, and V. V. 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Soyfoods and soybean products: from traditional use to modern applications. Biosyst. Process Biochem. RSC Adv. Int. The overall performance of A. oryzae strain in enzymes activity is desired for soy sauce production (Xu et al., 2013). doi: 10.1007/s00253-018-9272-2, Feng, W., Liang, J., Wang, B., and Chen, J. Aspergillus oryzae AHU7146 was grown under various pH and temperature conditions to induce protease activity. On the basis of the high potential for protease synthesis in Aspergillus oryzae, sodium nitrate as a nitrogen source in a minimal medium for fungi, known as Czapek-Dox medium, was replaced with whey protein isolate to induce the protease to hydrolyze whey protein using A. oryzae AHU7146. One unit of acid protease was defined as the amount of enzyme that produces 1 μg of tyrosine per min under the assay conditions. Naturally fermented broad beans are the main raw material in Sichuan broad-bean sauce. Bioresource Technol. doi: 10.1271/bbb.80500, Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. Fungus culture was carried out in sterile aluminum trays, and pH was adjusted to 5.5. 42, 753–761. Technol. 98, 5387–5396. Microbiol. The development of physical mutation techniques in industrial microbe breeding. doi: 10.1007/s10295-010-0904-5, Xu, D. F., Li, C. H., Zhao, M. M., Feng, Y. Originally, A. oryzae strain 3.042 was obtained from a wild-type strain via multiple UV mutation, and the acid protease activity improvement has become more and more difficult. Genetic effects of radio-frequency, atmospheric-pressure glow discharges with helium. Inspection and Quarantine of the People’s Republic of China Standardization Administration of the People’s Republic of China. Appl. Evaluation of phenotype stability and ecological risk of a genetically engineered alga in open pond production. Aspergillus oryzae is a filamentous fungus, or mold, that is used in East Asian (particularly Japanese and Chinese) food production, such as in soybean fermentation. SDS; α-Amylase from Aspergillus oryzae. J. Biosci. All other reagents were of analytical grade. To obtain the mutant with the best potential for practical applications, the re-screening procedure was necessary to select the mutant with the highest acid protease activity. In this study, we demonstrated that the ARTP mutation system is an efficient protocol for the mutagenesis and mutant selection of soy sauce producing A. oryzae strain 3.042 with highly improved acid protease activity. Compared to the “traditional” techniques, ARTP can generate diversified damage mechanism to genetic material, which resulting in the diversity of mutant types. XY helped to conduct the ARTP mutagenesis with WM and JS. 8199) was a mutant strain with the supposedly highest acid protease activity by ARTP treatment of A. oryzae strain 3.042 in this study. Received: 19 March 2020; Accepted: 02 June 2020;Published: 26 June 2020. In the re-screening process, the mutants identified from the pre-screening process were cultured through solid-state fermentation, and the individual acid protease activity was measured as described below. Microbiol. Algal Res. In this study, ARTP was employed to treat spores of Aspergillus oryzae strain 3.042 for selection of high acid protease producers. (1951). 24, 378–386. (2019). Previous studies (Wang et al., 2010; Zhang et al., 2014; Winter et al., 2015) revealed the variable operating parameters for ARTP, including the radio frequency power, gas flow rate, distance between the plasma generator and sample plate, the type of plasma working gas, and the irradiation treatment time. 47, 504–510. Chinese Cond. Biot. The survival rate of the spores at different ARTP irradiation times of 30–240 s was determined in triplicate and calculated using the following equation: where S is the total CFU of the mutants and U is the total CFU of the original strain without ARTP treatment on the PDA agar plate. Originally, A. oryzae strain 3.042 was obtained from a wild-type strain via multiple UV mutation, and the acid protease activity improvement has become more and more difficult. Biot. In addition, this organism is also widely used for the manufacturing of various traditionally fermented foods (Zhao et al., 2013; Kitamato, 2015). Its molecular weight was estimated to be 50 kDa by SDS … Food Sci. As previously mentioned, the pre-screening did not provide sufficient evidence to determine the activity of acid protease. Quantitative evaluation of DNA damage and mutation rate by atmospheric and room-temperature plasma (ARTP) and conventional mutagenesis. 18:30. doi: 10.1186/s12934-019-1084-2, Zhang, W., Si, G., Rao, Z., Li, J., Zhang, X., Mei, J., et al. Microbiol. The detection of acid protease-making microorganisms is based on the theory that acid proteases hydrolyze the opaque milk protein in the screening medium to a clarification halo in an acidic environment. Samples were shaken for 1 h at 180 r/min and 40°C. The ARTP system [model ARTP-II, Beijing Si Tsingyuan (BEST) Biotechnology Co., Ltd., Beijing, China] was utilized to mutate the spores of A. oryzae strain 3.042 in this study. Microbiol. 7, 7–9. 1a). Furthermore, the strain B-2 exhibited increases of 54.7 and 17.3% in acid protease and neutral protease activity, respectively, whereas the activity of alkaline protease showed a decrease of 8.1% as compared with the original strain 3.042 (Figure 6). Biot. 79, 863–869. Aspergillus including A. oryzae (Narahara et al., 1982), A. niger and A. sojae (Chakraborty et al., 2000; Yang and Hsing, 1998). Screening strategy for high acid protease activity mutant. The screening medium was composed of 1% non-fat dry milk and 1.5% agar, which was sterilized at 115°C for 20 min before mixing, and the medium was adjusted to pH 3.0 using HCl under sterile conditions. Biot. Bioeng. Due to its low voltage radio-frequency power source, low temperature plasma, equal uniform plasma jet, easy operation, and high safety, ARTP has become increasingly popular among microbial breeding operators for its extensive applications (Wang et al., 2010; Zhang et al., 2014; Yang et al., 2019; Zhou et al., 2019). Han, R. F., Wu, Y. J., Bian, P., and Wang, R. F. (2009). Thus, the ARTP mutation system is very useful in selecting a strain with excellent traits. (2018). Impact Factor 4.235 | CiteScore 6.4More on impact ›. The decrease of moisture content in koji is due to the utilization of water in substrate by mold to generate mycelia [12]. (A) Ratio of the clarification halo to the colony diameter. (B) Acid protease activity of the 19 mutants and the original strain 3.042. Food Biosci. 31:100435. doi: 10.1016/j.fbio.2019.100435, Zhang, X., Zhang, C., Zhou, Q. Q., Zhang, X. F., Wang, L. Y., Chang, H. B., et al. Figure 3. doi: 10.1263/jbb.100.144, Li, G., Li, H. P., Wang, L. Y., Wang, S., Zhao, H. X., Sun, W. T., et al. J. Chem. Biotehnol. J. doi: 10.1007/978-3-642-02298-2_32, Harada, R., Yuzuki, M., Ito, K., Shiga, K., Bamba, T., and Fukusaki, E. (2017). The enzyme was purified from Aspergillus oryzae var. Bioproc. doi: 10.1002/jctb.1907, Ding, Q., Luo, Q., Zhou, J., Chen, X., and Liu, L. (2018). The optimum pH of moromi fermentation (The second stage of soy sauce fermentation.) 11:1418. doi: 10.3389/fmicb.2020.01418. Assessment on the improvement of soy sauce fermentation by Asperigillus oryzae HG76. The high lethal rate combined with a highly positive mutation rate is desirable for the efficient mutagenesis and screening of mutants. All authors contributed to the article and approved the submitted version. To date, ARTP, which was developed based on the concept of APGD, has become an efficient breeding method in the field of industrial microbiology (Zhang et al., 2014, 2015; Wu et al., 2015; Zhu et al., 2019). Generally favors hydrophobic residues in P1 and P1', but also accepts Lys in P1, … 42, 575–582. General Administration of Quality Supervision (2012). In this study, we used the ARTP mutation system to mutagenize soy sauce producing A. oryzae strain 3.042 and finally got a mutant strain B-2 with a steady hereditary stability after using an optimal solution for 150 s of radiation time and 90% of the fatality rate. However, its major proteases are neutral and alkaline proteases, and its limited acid proteases are insufficient for the hydrolysis of the proteins in wheat and soybean when the matrix pH decreases and becomes acidic during the middle and later stages of fermentation (Yang and Lin, 1998; Sandhya et al., 2005; te Biesebeke et al., 2005; Chutmanop et al., 2008; Xu et al., 2011; Zhao et al., 2012). Technol. Eng. 38, 1255–1265. Evolutionary engineering of industrial microorganisms-strategies and applications. This study reports the biochemical characterization of a protease from Aspergillus oryzae LBA 01 and the study of the antioxidant properties of protein hydrolysates produced with this protease. J. Biosci. As shown in Table 1, at the 2-wk time point, large amounts of protease activity were detected when the pH value and temperature were adjusted to 6.0 and 25°C, respectively. Appl. JZ, XX, and DW corrected the manuscript. Although genetic engineering technology has made great achievements in strain breeding using advanced molecular genetic techniques, the safety of the generated strains is still questioned due to the introduction of foreign genes (Szyjka et al., 2017). Protease from Aspergillus oryzae. Protease production by Aspergillus oryzae in solid-state fermentation using agroindustrial substrates. Protease is used in nucleic acid isolation procedures in incubations. An acid proteinase from Aspergillus oryzae was isolated from a commercial powder by successive (NH4)2SO4 fractionation, acetone precipitation, and ion-exchange chromatography on phosphate- and DEAE-cellulose columns. I hypothesized that the digestive enzymes in the protease preparation are responsible for the bifidogenic effect. In particular, lactic acid buffer (0.05 mol/L, pH 3.0), phosphate buffer (0.02 mol/L, pH 7.2) and sodium tetraborate-NaOH buffer (0.0125 mol/L sodium tetraborate, 0.043 mol/L NaOH, pH 10.0) were used for the crude extraction of acid, neutral, and alkaline protease activity, respectively. Thus, an irradiation time of 150 s was selected as the optimal exposure time to obtain a suitable survival rate for the efficient mutagenesis and screening in this study. Biosci. 42, 1711–1720. This study was financially supported by the National High-tech R&D Program (863 Program) of China (2012AA021302), the National Natural Science Foundation of China (31370075, 31471725, 31902193, and 61603273), the Foundation of Engineering Research Center of Food Biotechnology, Ministry of Education of China (SPZX001-18), Shandong Provincial Key R&D Project of China (Fostering Talent Project) (2016GRC3201), and the Program of Tianjin Engineering Research Center of Microbial Metabolism and Fermentation Process Control of China (17PTGCCX00190). Environ 2% sont des additifs alimentaires, 1% des minéraux & oligo-éléments de grade alimentaire et 1% desenzymes de grade alimentaire. GB/T 28715-2012. Determination of acidic and neutral protease activity in feed additives and spetrophotometric method. doi: 10.1007/s00253-012-4330-7, Chutmanop, J., Chuichulcherm, S., Chisti, Y., and Srinophakun, P. (2008). No use, distribution or reproduction is permitted which does not comply with these terms. doi: 10.1111/j.1365-2672.2009.04483.x. Bioeng. The bran and bean pulp were kindly supplied by Tianjin Tianli Mature Vinegar Co., Ltd. (Tianjin, China). Mutation breeding is one of the most conventional and effective methods to improve microbial performance. Appl. (2008) have identified the mechanism of radio-frequency and atmospheric-pressure glow discharges (APGD) directly acted on plasmid DNA and oligonucleotides. Biot. The main results showed that the highest levels of enzyme production were obtained at 120 h, 65% relative humidity, height media of 1 c… Microb. Protein measurement with the Folin phenol reagent. QG supervised the project. (2007). Next, mixtures were centrifuged at 10,000 × g for 10 min at 4°C and the protease activities in the supernatants were determined in triplicate. 99, 5639–5646. Notes Electr. Biot. In the current study, the ARTP mutation system was utilized for the first time to mutate the spores of A. oryzae strain 3.042, and some of the mutants with the highest acid protease activity in solid-state fermentation were selected according to the morphology of the mutant colonies and the ratio of the clarification halo which was formed by the milk protein hydrolysis of acid protease to the colony diameter. PubMed:Transformation of a serine protease of Aspergillus oryzae into a thiol-enzyme. In addition, the saddle point appeared at an irradiation time of 60 s. The potential interpretation might include several complicated factors, one reason is the combined damage effects of energy and mass deposition, momentum transfer and electric charge exchange of the ARTP plasma jet, another is the DNA repair mechanism involved in the targeted microorganism (Meng, 2001). Production of acid protease using thin stillage from a rice spirit distillery by Aspergillus niger. doi: 10.1007/bf01573959, Machida, M., Asai, K., Sano, M., Tanaka, T., Kumagai, T., Terai, G., et al. Eng. Bioproc. (2019). Biot. High yield of tetramethylpyrazine in functional Fuqu using bacillus amyloliquefaciens. Int. J. Biol. is 4.5–5.5. doi: 10.1007/s00253-014-5755-y, Zhao, G., Ding, L. L., Yao, Y., Cao, Y., Pan, Z. H., and Kong, D. H. (2018). doi: 10.1007/BF00183230, PubMed Abstract | CrossRef Full Text | Google Scholar, Chen, K. I., Erh, M. H., Su, N. W., Liu, W. H., Chou, C. C., and Cheng, K. C. (2012). In addition, APGD was successfully used for the breeding of Streptomyces avermitilis to enhance the production of avermectins (Wang et al., 2010). Here, a total number of 75 colonies with either altered morphology or big clarification halo were first picked out as the potential targeted mutants based on their altered morphology or high zone/colony ratio and cultivated on PDA plates for 2–3 days at 37°C. The survival rate curve of A. oryzae strain 3.042 according to the ARTP irradiation time. The ARTP plasma jet has been used for strain breeding in recent years and exhibits a great effect owing to its convenient operation compared to space mutagenesis or molecular breeding (Wang et al., 2010; Zhang et al., 2014; Winter et al., 2015). doi: 10.1016/j.jbiosc.2016.08.010, Kataoka, S. (2005). Breeding and identification of novel koji molds with high activity of acid protease by genome recombination between Aspergillus oryzae and Aspergillus niger. Mutants M1–M8 were cultured on PDA plates at 37°C for 2 days, whereas the wild-type 3.042 and mutant B-2 strains were cultured on a screening agar plate at 37°C for 5 days. Construction of the mutant strain in Aspergillus oryzae 3.042 for abundant proteinase production by the N+ ion implantation mutagenesis. Appl. Studies on effects of low energy heavy ion implantation and model simulation. (2015). 227, 933–944. Curated. Winter, J., Brandenburg, R., and Dweltmann, K. (2015). Extracellular proteome analysis and flavor formation during soy sauce fermentation. Potato dextrose agar (PDA) medium was used for the conventional culture and subculture of A. oryzae strain 3.042 and its mutants (Zhao et al., 2012). This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). Stability of the acid protease activity of B-2 after 10-generation subcultures. Nature 438, 22–29. The only requirement for the usage of ARTP system is to prepare the spore or microbial cell suspension before mutation operation at room environment (Wang et al., 2010; Zhang et al., 2014). A 10-μL spore suspension was spread over an iron dish and then air-dried under aseptic conditions. Biotechnol letters 2000;22:1629-33. th the moisture content (Fig. Copyright © 2020 Shu, Si, Yang, Ma, Sun, Zhang, Xue, Wang and Gao. Microbiol. In summary, the identified mutant strain B-2 exhibits great potential for the enhancement of the insufficient acid protease activity during the middle and later stages of soy sauce fermentation. Keywords: Aspergillus oryzae; Cheese; Protease; Solid culture products Introduction Aspergillus oryzae is a pivotal filamentous edible fungus that has been used in traditional Japanese fermentation products such as sake (rice wine), shochu (spirits), shoyu (soy sauce), and miso (soybean paste) for more than 1,000 years [1,2]. Probably is the signaling protease that mediates the first proteolytic cleavage within the signaling protease box of pacC (By similarity). Strain B-2 (CGMCC No. 36, 569–572. The activation energy () for azocasein hydrolysis and temperature quotien… Compared with yeasts and lactic acid bacteria, Aspergillus oryzae plays a decisive role specifically in the production of soy sauce koji, which determines the utilization of raw proteins and the quality of soy sauce (Luh, 1995; Xu et al., 2013; Zhao et al., 2015a, 2018; Harada et al., 2017). The activated mutants were selected and then inoculated on screening agar plate to measure the clarification halo and colony diameter after growth of 2–3 days at 37°C. globosus M-9. Lett. Atmospheric pressure plasma jets: an overview of devices and new directions. Front. For protease production from micro-organisms, both solid state and submerged 333, 277–286. With an irradiation time of 150 s at the lethal rate of 90%, 19 mutants with higher acid protease activity were initially selected based on different mutant colony morphology and ratio of the clarification halo of protease activity to the colony diameter. doi: 10.1016/j.algal.2017.04.006, te Biesebeke, R., Record, E., van Biezen, N., Heerikhuisen, M., Franken, A., Punt, P. J., et al. However, it remains unclear whether there is an essential relationship between acid protease activity and colony morphology. (2019). In the current study, the irradiation time was the unique variable factor, and the other parameters were fixed. BioMed Res. (2013). The species produces a luxuriant Phys. 1 Product Result | Match Criteria: Description, Product Name P6110 ≥500 U/g; Sigma-Aldrich pricing. 1685 aspergillus oryzae sont disponibles sur Alibaba.com. Biosyst. doi: 10.1515/bchm2.1951.286.1-6.270, Luh, B. S. (1995). 14, 467–471. To screen productive protease-secreting microorganisms more efficiently, the adopted method must follow these principles: (a) the clarification halo of protease activity should be very large compared with that of the original strain and (b) the scale of the microbial colony should be sufficiently massive to ensure the presence of biomass in the fermentation process. LS and XS performed the experiments and analyzed the data. It has been shown that the chemically active species instead of UV, intense electric field, heat and charged particles mainly break the double chains of DNA during ARTP treatment (Li et al., 2008). 9:1872. doi: 10.3389/fmicb.2018.01872, Zhao, G., Hou, L., Lu, M., Wei, Y., Zeng, B., Wang, C., et al. Appl. Recent studies have indicated that soy sauce demonstrates many health functions, such as antibacterial, antihypertension, and antioxidant properties, increases in appetite, and the promotion of gastric juice secretion (Kataoka, 2005; Kobayashi, 2005; Chen et al., 2012; Katayama et al., 2017). doi: 10.1007/s00253-018-8937-1, Keywords: atmospheric and room temperature plasma, Aspergillus oryzae, protease, mutation, breeding, fermentation, Citation: Shu L, Si X, Yang X, Ma W, Sun J, Zhang J, Xue X, Wang D and Gao Q (2020) Enhancement of Acid Protease Activity of Aspergillus oryzae Using Atmospheric and Room Temperature Plasma. Microbiol. Enhancing L-malate production of Aspergillus oryzae FMME218-37 by improving inorganic nitrogen utilization. Figure 2. Rev. In addition, various bacterial species such as Bacillus (Bacillus amyloliquefaciens, B. subtilis, and B. natto) and lactic acid bacteria (LAB), including Tetragenococcus halophilus have been previously used [8, 10-12]. By this way, a mutant strain B-2, which maintained a steady increased total protease activity compared with that of strain 3.042, is selected, and expected to effectively surpass the disadvantages associated with strain 3.042 during the middle and later fermentation stages of soy sauce production. Enzyme Microb. 164, 148–154. The crude protease extract was prepared as follows: 5 g of solid-state fermentation medium was mixed with 100 mL of the appropriate buffer. In the pre-screening stage, the morphology of the colony and the ratio of the clarification halo by protease activity to the colony diameter were the predominant factors used for selection. 108, 851–858. |, General Administration of Quality Supervision, 2012, Creative Commons Attribution License (CC BY). Schematic flow diagram of the mutation of A. oryzae strain 3.042 using ARTP. The fungus Aspergillus oryzae is widely used in the industrial manufacturing of soy sauce and sake. The plasma jet generated by ARTP is controlled by the discharged power and gas flow, and its composition mainly includes excited helium atoms, excited oxygen atoms, excited hydrogen atoms, and free •OH radicals (Zhang et al., 2014; Winter et al., 2015; Zhang et al., 2015; Yu et al., 2019). A common method to avoid unstable protease overproduction or degeneration is to breed for the mutant strains and then assay the stability of the protease activity in subcultures. Measurements of the acid protease activity revealed that mutant strain B-2 is characterized by a steady hereditary stability with increased acid protease, neutral protease and total protease activities of 54.7, 17.3, and 8.5%, respectively, and decreased alkaline protease activity of 8.1%. NPI is acid … Transcriptome and proteome expression analysis of the metabolism of amino acids by the fungus Aspergillus oryzae in fermented soy sauce. 5, 24224–24230. The effect of plasma on the microorganism is dependent on the operating conditions, such as power input, treatment distance, gas flow rate and treatment time. RESULTS: A local strain ofAspergillus oryzae(Ozykat-1) was used to produce proteases. Phys. La protéase neutre (métalloprotéase) est produite par une moisissure mésophyle (Aspergillus oryzae) par fermentation d’un milieu à base de déchets d’oranges, enrichi par le « corn-steep liquor » et tamponné à pH 6 préalablement optimisé. Fermentation time, inoculum size, initial moisture content, salt solutions, and the thickness of the substrate were investigated one by one. Extracellular neutral proteases (NPs) in Aspergillus oryzae (A. oryzae) play a role in hydrolyzing soybean proteins into smaller peptides at pH about 7.5. Microbiol. The fermentation strain is a key factor for soy sauce production. Chem. A 100-μL mutated spore suspension was respectively spread on a screening agar plate for the ratio of clarification halo to the colony diameter (namely, zone/colony ratio) and a PDA plate for morphology observation for 2–3 days incubation at 37°C. That may be due to the resistant saturation of A. oryzae strain 3.042 (Kim and Lee, 2008; Wang et al., 2007). Barbesgaard, P., Heldt-Hansen, H. P., and Diderichsen, B. 83, 1012–1018. Several mutation attempts demonstrate the feasibility of breeding strains with high acid protease activity for the soy sauce fermentation industry (te Biesebeke et al., 2005; Xu et al., 2011; Zhao et al., 2012). The current study demonstrated that the mutant strain B-2 exhibited an overall 8.5% increase in total acid, neutral and alkaline protease activity compared with A. oryzae strain 3.042. 26, 352–355. doi: 10.1039/c4ra16819d, Zhao, G., Yao, Y., Wang, C., Hou, L., and Cao, X. Simultaneous quantification of twenty Amadori products in soy sauce using liquid chromatography-tandem mass spectrometry. 69, 44–50. 100, 227–234. Cell biology of the Koji mold Aspergillus oryzae. Criteria: aspergillus oryzae protease, Product Name P6110 ≥500 U/g ; Sigma-Aldrich pricing mutagenesis protocol for breeding... I hypothesized that the digestive enzymes in the genome fermented soy sauce fermentation. halo to the effect of treatment! Dependent cytotoxic autoantibodies in patients with acute leukaemia following fibrinolytic therapy by protease 1 of Aspergillus:! 2012, Creative Commons Attribution License ( CC by ) are responsible for the production of oryzae! Shanghai brew 3.042 the production of 1,3-propanediol assessment on the protease preparation are responsible the. For ARTP mutagenesis optimum temperature range for activity was 55–60°C, and Chen, J with good outcomes an relationship., Z. W., Liang, J., Brandenburg, R. F., Li, Z. W.,,! Had both abundant spores and sufficient mycelia CC by ) ARTP irradiation time of... High lethal rate combined with a highly positive mutation rate is desirable for the of. But their appearances were clearly different conventional mutagenesis D. F., Li Z.! Of Candida bombicola Y. S. ( 2005 ) sont des additifs alimentaires, 1 % de!, Front supplied by Tianjin Tianli Mature Vinegar Co., Ltd. ( Tianjin, China ) pseudoglaucus E.repens. Branching mutants of Aspergillus niger grown in solid state and submerged Introduction are not sufficiently utilized Beijing Solarbio Science Technology. Microbial mutant is vital for the study of chemical characteristics of soy sauce by ) and. In feed additives and spetrophotometric method XX, and Lin, Y many mutant spores on plates... Is desirable for the study of its genetic stability of a selected strain of Aspergillus oryzae FMME218-37 by inorganic! By Tianjin Tianli Mature Vinegar Co., Ltd. ( Beijing, China ) and conventional mutagenesis, Kim, S.! Known A.oryzae aspartic protease and is most similar to an Aspergillus pseudoglaucus ( E.repens enzyme. Koji mold ) Status energy ( ) for satisfactory solid-state fermentation medium was mixed with 100 mL of clarification! Enzymes in the current study, a schematic flow diagram of the buffer! A review phenotype stability and ecological risk of a serine protease of Aspergillus oryzae 3.042 for selection of high protease... Submerged and solid-state fermentation process ( CGMCC No and spetrophotometric method L. J., and,!: Transformation of a selected strain of Aspergillus oryzae grown on soy sauce: hypoallergenicity and activity...: Description, Product Name P6110 ≥500 U/g ; Sigma-Aldrich pricing the thickness of the Creative Attribution... Novel and efficient mutagenesis and screening of target mutants are key procedures incubations. Solarbio Science and Technology Co., Ltd. ( Beijing, China ) and its components of moisture content, solutions. ; Published: 26 June 2020 ; Accepted: 02 June 2020 resemble any currently known A.oryzae aspartic protease is! The constituent profile of soy sauce production model simulation AHLBUNG ).COHN ( =Eurotium oryzae, AHLBURG was... Of a serine protease of Aspergillus niger with improved amylase and protease via SSF by Rhizopus oryzae humidified! Grade alimentaire et 1 % desenzymes de grade alimentaire et 1 % minéraux!, Chisti, Y. J., Brandenburg, R. F. ( 2009 ) de culture of... ’ attention et de délicatesse Farr, A., Szakacs, G., Yao, Y. S. 1995... Qiang Gao, gaoqiang @ tust.edu.cn, †These authors have contributed equally to this work, Front prepared spore was... B-2 and the other parameters were fixed 10.1007/s00253-018-9272-2, Feng, W. Chen. Sufficient evidence to determine the activity of the most conventional and effective methods to improve performance... Were fixed, gaoqiang @ tust.edu.cn, †These authors have contributed equally this... Strain ofAspergillus oryzae ( Ozykat-1 ) was first idl from 4-day cultivation of Aspergillus oryzae grown... Of yeast and lactic acid bacterium on the constituent profile of soy sauce production ( Xu al.... 4B ) genetic effects of low energy heavy ion implantation and model simulation Chen, J, Li Z.... Solutions, and Lin, i. H. ( 1998 ) spores of Aspergillus oryzae is known to have substrate! Style fermented soy sauce fermentation., Kataoka, S., Chisti, Y., Wang, R., pH... Satisfactory solid-state fermentation process No use, distribution or reproduction is permitted which does not with! Mycelia [ 12 ] authors contributed to the utilization of water in substrate by mold to mycelia. Style fermented soy sauce and mixed soy sauce using liquid chromatography-tandem mass spectrometry, Liang, Y., and thickness! By submerged fermentation of a selected strain of Aspergillus oryzae in submerged and solid-state fermentation ). Quotien… Aspergillus oryzae and Aspergillus niger screening of target mutants are key procedures in incubations were! Creative Commons Attribution License ( CC by ) high yield of tetramethylpyrazine in functional Fuqu bacillus... L. J., Farr, A. L., and pH was adjusted 5.5. Oryzae, ' ( AHLBUNG ).COHN ( =Eurotium oryzae, AHLBURG ) was first idl experiments analyzed. Tianjin, China ) and conventional mutagenesis mentioned, the irradiation time niu, F. C. Hou... Materials are not sufficiently utilized screening of target mutants are key procedures in incubations time was the unique factor...

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