Protease Production by Different Thermophilic Fungi

Macchione, Mariana M.; Merheb, Carolina W.; Gomes, Eleni; Silva, Roberto da

doi:10.1007/978-1-60327-526-2_34

Cited by 7 publications

(8 citation statements)

References 14 publications

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“…The alkaline serine protease gene ALP5 was successfully expressed in P. pastoris and, as expected, the proteolytic activity of the recombinant protease showed its highest activity under alkaline conditions (at pH 10) and temperature of 50 °C, which was very similar to the activity of the recombinant protease expressed in E. coli and to other alkaline serine proteases of bacterial (Gupta et al, 2002) and fungal (Macchione et al, 2008) origin. These results are consistent with a previous report indicating that the optimal pH and temperature of a related protein (alkaline α-amylase gene) from alkaliphilic Alkalimonas amylolytica expressed both in E. coli and P. pastoris are similar (Yang et al, 2012).…”

Section: Discussionsupporting

confidence: 69%

Biocontrol activity of an alkaline serine protease from Aureobasidium pullulans expressed in Pichia pastoris against four postharvest pathogens on apple

Banani

Spadaro

Zhang

et al. 2014

International Journal of Food Microbiology

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38The yeast-like fungus Aureobasidium pullulans PL5 is a microbial antagonist against postharvest 39 pathogens of fruits. The strain is able to produce hydrolases, including glucanases, chitinases and 40 proteases. The alkaline serine protease gene ALP5 from A. pullulans was cloned, inserted into the 41 vector pPIC9 to construct pPIC9/PL5, and then expressed in Pichia pastoris strain KM71. ALP5 42 had a molecular mass of 42.9 kDa after 5 days growth with 1% methanol induction at 28 °C. The storage technologies are available (Khamis et al., 2012; Martini et al., 2013; Snowdon, 1990). To 78Among the different biocontrol agents, yeasts are promising and gaining popularity (Jamalizadeh et 79 al., 2011; Janisiewicz et al., 2010; Spadaro et al., 2008). In particular, the yeast-like fungus 80Aureobasidium pullulans De Bary (Arnaud), showed to be effective against B. cinerea, P. 81expansum and Rhizopus stolonifer on various fruit, including apple, grapes, sweet cherry, 82 strawberry and peach (Bencheqroun et al., 2007; Ippolito et al., 2000; Lima et al., 1997; Schena et 83 al., 2003). Moreover, the strain PL5 of A. pullulans showed high efficacy in the control of B.84 cinerea and P. expansum on apples, in addition to Monilinia laxa on plums and peaches (Zhang et 85 al., 2010a). 86Several mechanisms have been reported to play a significant role in the biocontrol activity of A. 87pullulans strains, including induction of defence responses (Ippolito et al., 2000) and competition 88for nutrients (Bencheqroun et al., 2007). Recently, it was demonstrated that the strain PL5 secretes protease (Zhang et al., 2010a; 2012). 91In mycoparasitism, fungal proteases may be significantly involved in antagonistic activity, because 92 they may play a significant role in fungal cell wall lysis, which is composed of chitin and glucan 93 polymers embedded in, and covalently linked to a protein matrix (Wessels, 1986 Construction of the expression vector pPIC9-ALP5 156The product was subsequently digested with EcoRI and ligated into the pPIC9 vector, which was The pPIC9-ALP5 construct was linearized by Stu I enzyme for efficient integration into the P. 165pastoris genome, then it was transformed into P. pastoris strain KM71. were done to confirm whether the protease cDNA was integrated into the genomic DNA of P. 171pastoris. Primers used for PCR, 5'AOX1 (5'-GACTGGTTCCAATTGACAGC-3') and 3'AOX1 172(5'-GCAAATGGCATTCTGACATCC-3') were provided by the manufacturer. DC and DP refers to the average diameters of fungal mycelia of control and protease respectively. 227The experiment was performed three times. Results 271 Expression and purification of recombinant protease 272The expression of purified recombinant protease from a transformed isolate of P. pastoris was 273 analysed through SDS-PAGE (Figure 1). After 120 h induction, the protease band was observed in 274 some isolates at 42.9 kDa, which corresponds to the same molecular weight of the ALP5 protease selected for large-scale expression. The recombinant protein was e...

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Section: Discussionsupporting

confidence: 69%

Biocontrol activity of an alkaline serine protease from Aureobasidium pullulans expressed in Pichia pastoris against four postharvest pathogens on apple

Banani

Spadaro

Zhang

et al. 2014

International Journal of Food Microbiology

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“…The study of extremophiles allows us to discover and recognize more and more ways of adaptation of organisms and proteins, which gives a possibility to approach individualized proteins designed for various stressful conditions. Such protein-enzymes can help in solving many problems, in particular, waste disposal, water purification, and biofuel production [15,16].…”

Section: F I G 1 Keratinase Activity Of Thermophilic Fungi On the 14 Th Day Of Cultivationmentioning

confidence: 99%

“…We have shown a high keratinase activity of C. thermophilum, R. tauricus, T. thermophilus, T. terrestris, T. auranticus (10-26 U/mL). Strains of these species have repeatedly acted as producers of proteolytic enzymes [16,17,33], although they were an order of magnitude inferior activity to representatives of Aspergillus terreus [34]. The high keratinase activity of studied thermophilic fungi testifies to the high potential of this group for the search for producers of keratinolytic enzymes, which are in demand by the processing and feed industry.…”

Section: F I G 4 β-Mannanase and Cm-cellulase Activity Of Thermophilic Micromycetes (6 Days Of Cultivation 42 °C)mentioning

confidence: 99%

Thermophilic Fungi with Glucosidase and Proteolytic Activities

Borzova¹,

Гудзенко²,

Avdiyuk³

et al. 2021

Mikrobiol. Z.

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The directed search for extremophilic producers in order to obtain hydrolytic enzymes with increased thermal stability has an unconditional practical potential for use in the food and feed industry to improve the quality of the final product. The aim of the work was to study the ability of collection strains of thermophilic fungi to show α-L-rhamnosidase, α-galactosidase, cellulase, β-mannanase, keratinase and caseinolytic activity. Methods. Micromycetes were grown under submerged conditions in test tubes at 42°C for 8–14 days. Enzymatic activities were studied in the culture liquid supernatant. p-Nitrophenyl-α-D-galactopyranoside, naringin, guar gum galactomannan and Na-carboxymethylcellulose were used as substrates to determine α-galactosidase, α-L-rhamnosidase, β-mannanase and cellulase activities, respectively. Casein and crushed defatted feathers were served as substrates for the determination of proteolytic activity. Results. The enzymatic activity of 50 strains of micromycetes belonging to 17 species was investigated. The studied group showed high activity: 94% of the strains had at least one, 34% – two, 26% – from three to five enzyme activities. The most active keratinase producers were Thielavia terrestris 1920 and 62, Rhizomucor tauricus 1909, Chrysosporium thermophilum 2050, Thermoascus thermophilus 92 and Thermoascus aurantiаcus 2052 (10–26 U/mL). The highest α-L-rhamnosidase activity was observed in T. terrestris 62 (0.35 U/mL), and carboxymethylcellulase activity −in Thermomyces lanuginosus 2046. Six strains showed α-galactosidase (0.05–0.2 U/mL) and four strains − β-mannanase (5–130 U/mL) activity. Conclusions. As a result new strains producing proteolytic and glycolytic enzymes were isolated among thermophilic micromycetes. Soil thermophilic micromycetes can be used as producers of proteolytic and glycolytic enzymes. Of particular interest are the cultures of Acremonium thermophilum 1963, Corynascus thermophilum 2050, C. sepedonium 1899 and 65068, T. thermophilus 1946, which are capable of producing complexes of proteases and glycosidases in the culture liquid. This indicates that these strains are promising for use as destructors in various technologies processing of complex raw materials.

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“…Some workers 5,17,29 also reported the similar investigations. Macchione et al 23 worked on protease production by different thermophilic fungi and reported that wheat bran was best substrate for protease production in solid state fermentation and soybean meal extract was best substrate for protease production in submerged fermentation. These differences in findings might be due to different nature of fungus and different composition of substrate favored for protease production.…”

Section: Screening Of Substratesmentioning

confidence: 99%

Exploitation of Different Agro-residues for Acid protease Production by Rhizopus sp. in Koji Fermentation

Irfan¹,

Rauf²,

Syed³

et al. 2011

IJAVMS

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Two different strains of Rhizopus sp. i.e. R. oligosporus-M30 and R. arrhizus-M26 were employed for the production of acid protease using different agro-residues such as rice husk, rice bran, wheat bran, defatted soybean meal and sunflower meal as a sole carbon source. R. oligosporus-M30 gave maximum enzyme level with soybean meal (15.6 U/gds) while R. arrhizus-M26 with sunflower meal (13.6 U/gds) respectively. The optimal conditions for the enzyme production by R .oligosporus-M30 were initial pH of 5.5, temperature 30 o C with 10% of inoculum size for 72hr of fermentation period. When R. arrhizus-M26 was cultivated that exhibit the best enzyme production with the fermentation period of 72h, initial pH 5.0, temperature 33 o C with 5% of inoculum size

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Protease Production by Different Thermophilic Fungi

Cited by 7 publications

References 14 publications

Biocontrol activity of an alkaline serine protease from Aureobasidium pullulans expressed in Pichia pastoris against four postharvest pathogens on apple

Biocontrol activity of an alkaline serine protease from Aureobasidium pullulans expressed in Pichia pastoris against four postharvest pathogens on apple

Thermophilic Fungi with Glucosidase and Proteolytic Activities

Exploitation of Different Agro-residues for Acid protease Production by Rhizopus sp. in Koji Fermentation

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