RESUMO
Background: To identify the critical amino acid residues that contribute to the high enzyme activity and good thermostability of Yersinia enterocolitica subsp. palearctica (Y. NSN), 15 mutants of Y. NSN were obtained by site-directed mutagenesis in this study. And their enzyme activity and thermostability were assayed. Effect of several factors on the enzyme activity and thermostability of Y. NSN, was also investigated. Results: The results showed that the I203F and D264E mutants retained approximately 75% and 70% enzyme activity, respectively, compared to the wild-type enzyme. In addition to the I203F and D264E mutants, the mutant E202A had an obvious influence on the thermostability of Y. NSN. According to the analysis of enzyme activity and thermostability of Y. NSN, we found that Glu202, Ile203 and Asp264 might be the key residues for its high enzyme activity and good thermostability. Conclusions: Among all factors affecting enzyme activity and thermostability of Y. NSN, they failed to explain the experimental results well. One reason might be that the enzyme activity and thermostability of Y. NSN were affected not only by a single factor but also by the entire environment.
Assuntos
Desoxirribonucleases/química , Desoxirribonucleases/genética , Yersinia enterocolitica/enzimologia , Endonucleases/química , Endonucleases/genética , Ensaios Enzimáticos , Estabilidade Enzimática , Temperatura Alta , Mutagênese Sítio-DirigidaRESUMO
ABSTRACTIn this work, the mixture of alginate and soy protein isolate used as a wall material was developed to encapsulateEnterococcus faecalis HZNU P2 (E. faecalis HZNU P2). The survival ability in the simulated gastric fluid (SGF) and bile salt solution, storage stability at different temperatures and release properties in the simulated intestinal fluid (SIF) of encapsulated cells were assessed. The results showed that encapsulation could offer sufficient protection toE. faecalis HZNU P2. The viability of encapsulatedE. faecalis HZNU P2 did not decrease in SGF at pH 2.5 or 2.0 after 2 h incubation, while free cells were reduced from 11 to 9.85 log CFU/mL in SGF (pH 2.5) at the same exposure time. Only minor viability of encapsulatedE. faecalis HZNU P2 lost in 1.0 or 2.0% bile salt solution for 1 or 2 h exposure, compared with no survival of freeE. faecalis HZNU P2 under the same conditions. EncapsulatedE. faecalis HZNU P2 was completely released from the microspheres in SIF within 1 h. The viability of encapsulatedE. faecalisHZNU P2 stored for two weeks at 4°C was fully retained. Viabilities of encapsulatedE. faecalis HZNU P2, 9.6 and 9.0 Log CFU/g were obtained at 25 and 37°C after 21 days storage, respectively. However, around 1.0 log CFU/mL of free cells was reduced after two weeks storage at 4°C. EncapsulatedE. faecalis HZNU P2 using soy protein isolate and alginate as wall materials could play an important role in food applications.
RESUMO
Date is the most popular fruit in middle-east countries. However, the date production process is accompanied by a substantial increase of loss during picking, storage, commercialization and conditioning process. Date and their byproducts have many essential elements for the growth of microorganisms. Thus, they can be converted into value-added compounds through biotechnology. In this paper, date and their processing byproducts used as substrates for producing value-added products such as organic acids, exopolysaccharide, antibiotics, date flavored probiotic fermented dairy, bakery yeast, etc, were reviewed.
RESUMO
In this work, alginate-whey protein was used as wall materials for encapsulating Lactobacillus delbrueckii subsp. bulgaricus (L. bulgaricus). The characteristics of encapsulated and free L. bulgaricus showed that the free L. bulgaricus lost viability after 1 min exposure to simulated gastric fluid (SGF) at pH 2.0 and 2.5. However, the viability of encapsulated L. bulgaricus did not decrease in SGF at pH 2.5 for 2 h incubation. The viable numbers of encapsulated L. bulgaricus decreased less than 1.0 log unit for 2 h incubation in SGF at pH 2.0. For bile stability, only 1.2 log units and 2.0 log units viability of the encapsulated L. bulgaricus was lost in 1 and 2% bile for 1 h exposure, respectively, compared with no survival of free L. bulgaricus under the same conditions. Encapsulated L. bulgaricus was completely released from the microspheres in simulated intestinal fluid (SIF, pH 6.8) in 3 h. The viability of the encapsulated L. bulgaricus retained more 8.0 log CFU/g after stored at 4°C for four weeks. However, for free L. bulgaricus, only around 3.0 log CFU/mL was found at the same storage conditions. Results showed that the encapsulation could improve the stability of L. bulgaricus.