来源于Cronobacter universalis的植酸酶酶学性质的研究

按照毕赤酵母(Pichia pastoris)对密码子的选择偏向性,对来源于细菌(Cronobacter universalis)的植酸酶基因(以下命名为CuPhyS)进行了密码子优化改造,合成并且作为外分泌蛋白成功地在毕赤酵母GS115里表达,CuPhyS的蛋白分泌量可积累到45μg/mL。酶学性质研究表明:该酶的最适pH为4.0,最适温度为50℃,在pH 3.0—12.0时该酶比较稳定,在50℃下热稳定性较高;以植酸钠作为底物,Mg^(2+)对酶活有一定的激活作用,Cu^(2+)对酶活有一定的抑制作用;该酶还具有良好的抗胰蛋白酶水解的能力和部分抗胃蛋白酶水解的能力。

Insights into Cronobacter sakazakii Biofilm Formation and Control Strategies in the Food Industry

Cronobacter sakazakii(C.sakazakii)is a foodborne opportunistic pathogen that can cause life-threatening invasive diseases,such as necrotizing enterocolitis,meningitis,and sepsis in infants.The potential risk of C.sakazakii contamination of powdered infant formula(PIF)is an issue that has attracted considerable attention from manufacturers,regulators,and consumers.C.sakazakii biofilms on the surfaces of equipment and in diverse food-production environments constitute a mode of cell growth that protects the pathogen from hostile environments,and are an important source of persistent contamination of food products.Bacterial biofilms are difficult to remove due to their resistant properties.Conventional cleaning and sterilizing procedures may be insufficient for biofilm control,and may lead to further biofilm development and dispersal.Consequently,novel control strategies are being developed,such as nanotechnology-based delivery systems,interspecies interactions,antimicrobial molecules of microbial origin,natural extracts,and phages.This review focuses on describing the mechanisms underlying the biofilm formation and behavior of C.sakazakii and discussing potential control strategies.

Food Safety Risks and Contributing Factors of Cronobacter spp.

Cronobacter species are a group of Gram-negative opportunistic pathogens,which cause meningitis,sep-ticemia,and necrotizing enterocolitis in neonates and infants,with neurological sequelae in severe cases.Interest in Cronobacter has increased significantly in recent years due to its high virulence in children.In this review,we summarize the current understanding of the prevalence of Cronobacter species in several important food types.We discuss the response mechanisms enabling persistence in adverse growth con-ditions,as well as its pathogenicity.We emphasize the food safety concerns caused by Cronobacter and subsequent control methods and clinical treatments.

Application of GFPuv Labeled Cronobacter sakazakii for Evaluation of Its Survival during Cornstarch Processing

Cronobacter sakazakii is an emerging pathogen that can cause diseases for several infant groups. These bacteria were contaminated in foods, clinical utensils, and environments. In Indonesia, C. sakazakii has been isolated from powdered infant formulas, weaning foods, and other dried foods such as cornstarch. The objective of this research is to trace survival of C. sakazakii during cornstarch production step using its mutant. Mutant was constructed by inserting Green Fluorescent Protein plasmid inside to the bacterial cell that appeared green fluorescent colonies under UV observation. The presence of C. sakazakii during processing was conducted by artificial contamination. This research consists of three steps, i.e. determination of the suitable enumeration method of C. sakazakii’s mutant, cornstarch production from yellow corn, and survival analysis of C. sakazakii during endosperm soaking and cornstarch drying. The suitable enumeration method was surface plating method on TSA-ampicillin medium combining with UV light application because of ineffectiveness of ampicillin inhibition for growth of yeasts and molds. The cornstarch produced in laboratory has the same properties with commercial cornstarch in parameters of moisture content, density, and starch granule structure. The yield of cornstarch final product was 48.90% (dry whole kernel-based). C. sakazakii cannot survive in 48 hours soaking process at 52?C and 24 hours drying process at 50?C that is applied during cornstarch production.

Cronobacter新属(阪崎肠杆菌)检测分型方法的研究进展

本文对Cronobacter新属的以VRBG琼脂、NA+α-MUG、DFI、ESIA或ESPM等选择性分离培养基进行分离培养的传统培养方法和基于16SrRNA序列、基因组重复序列、16S-23SrDNA ITS、tRNA-glu和23SrDNA之间DNA序列、gluA、rpsU与dnaG及ompA等目标区域的分子生物学检测分型方法进行了综述。

Cronobacter spp.,foodborne pathogens threatening neonates and infants

Cronobacter spp.(formerly Enterobacter sakazakii) are special foodborne pathogens. Cronobacter infection can cause necrotizing enterocolitis, sepsis and meningitis in all age groups, especially neonates and infants, with a high fatality of up to 80%, although the infection is rare. Outbreaks of Cronobacter infection are epidemiologically proven to be associated with contaminated powdered infant formula(PIF). Cronobacter spp.can resist dry environments and survive for a long period in food with low water activity. Therefore, Cronobacter spp.have become serious pathogens of neonates and infants, as well as in the dairy industry. In this review, we present the taxonomy, pathogenesis, resistance, detection and control of Cronobacter spp.

Ketogenic diet poses a significant effect on imbalanced gut microbiota in infants with refractory epilepsy

AIM To investigate whether patients with refractory epilepsy and healthy infants differ in gut microbiota(GM),and how ketogenic diet(KD) alters GM.METHODS A total of 14 epileptic and 30 healthy infants were recruited and seizure frequencies were recorded. Stool samples were collected for 16 S r DNA sequencing using the Illumina Miseq platform. The composition of GM in each sample was analyzed with MOTHUR,and intergroup comparison was conducted by R software.RESULTS After being on KD treatment for a week,64% of epileptic infants showed an obvious improvement,with a 50% decrease in seizure frequency. GM structure in epileptic infants(P1 group) differed dramatically from that in healthy infants(Health group). Proteobacteria,which had accumulated significantly in the P1 group,decreased dramatically after KD treatment(P2 group). Cronobacter predominated in the P1 group and remained at a low level both in the Health and P2 groups. Bacteroides increased significantly in the P2 group,in which Prevotella and Bifidobacterium also grew in numbers and kept increasing.CONCLUSION GM pattern in healthy infants differed dramatically from that of the epileptic group. KD could significantly modify symptoms of epilepsy and reshape the GM of epileptic infants.

Characterization and Optimization of Amylase Producing Bacteria Isolated from Solid Waste

Municipal waste is one of the most hazardous components of developing countries. However, enzymes do provide an eco-friendly solution in this case. Amylase is an important enzyme in food, textile and pharmaceutical industry and can be used for bioconversion of waste. From the municipal solid waste we have isolated an amylase producing bacteria that can grow in the irritant municipal waste and help in their bio conversation. The bacteria were identified as Cronobacter sakazakii Jor52 (C2). The optimized media for maximum amylase production after 24 h of incubation, contains 2% starch, 0.6% peptone, 0.01% CaCl2, 0.05% KCl, 0.05% MgSO4 and 0.05% K2HPO4. The crude enzyme activity and stability study revealed that the amylase is stable within the pH 6 - 8 and temperature 30°C - 40°C and give maximum activity at 37°C at pH-8.

Unusual metallo-β-lactamases may constitute a new subgroup in this family of enzymes

Metallo-β-lactamases (MBLs) are a family of Zn2+-dependent enzymes that have contributed strongly to the emergence and spread of antibiotic resistance. Novel members as well as variants of existing members of this family are discovered continuously, compounding their threat to global health care. MBLs are divided into three subgroups, i.e. B1, B2 and B3. The recent discovery of an unusual MBL from Serratia proteamaculans (SPR-1) suggests the presence of an additional subgroup, i.e. B4. A database search reveals that SPR-1 has only one homologue from Cronobacter sakazakii, CSA-1.These two MBLs have a unique active site and may employ a mechanism distinct from other MBLs, but reminiscent of some organophosphate-degrading hydrolases.