Safety assessment of a novel marine multi-stress-tolerant yeast Meyerozyma guilliermondii GXDK6 according to phenotype and whole genome-sequencing analysis

The application of microorganisms as probiotics is limited due to lack of safety evaluation.Here,a novel multi-stress-tolerant yeast Meyerozyma guilliermondii GXDK6 with aroma-producing properties was identified from marine mangrove microorganisms.Its safety and probiotic properties were assessed in accordance with phenotype and whole-genome sequencing analysis.Results showed that the genes and phenotypic expression of related virulence,antibiotic resistance and retroelement were rarely found.Hyphal morphogenesis genes(SIT4,HOG1,SPA2,ERK1,ICL1,CST20,HSP104,TPS1,and RHO1)and phospholipase secretion gene(VPS4)were annotated.True hyphae and phospholipase were absent.Only one retroelement(Tad1-65_BG)was found.Major biogenic amines(BAs)encoding genes were absent,except for spermidine synthase(JA9_002594),spermine synthase(JA9_004690),and tyrosine decarboxylase(inx).The production of single BAs and total BAs was far below the food-defined thresholds.GXDK6 had no resistance to common antifungal drugs.Virulence enzymes,such as gelatinase,DNase,hemolytic,lecithinase,and thrombin were absent.Acute toxicity test with mice demonstrated that GXDK6 is safe.GXDK6 has a good reproduction ability in the simulation gastrointestinal tract.GXDK6 also has a strong antioxidant ability,β-glucosidase,and inulinase activity.To sum up,GXDK6 is considered as a safe probiotic for human consumption and food fermentation.

Changes of composition and antibiotic resistance of fecal coliform bacteria in municipal wastewater treatment plant

The dynamics of the composition and antibiotic resistance of the fecal coliform bacteria(FCB)in a typical wastewater treatment plant(WWTP)were investigated concerning the seasonal changes.Results showed that WWTP could remove the FCB concentration by 3∼5 logs within the effluent of 10^(4)∼10^(5)CFU/L,but the antibiotic resistant rate of FCB species increased significantly after WWTP.The dominant FCB changed from Escherichia coli in the influent(∼73.0%)to Klebsiella pneumoniae in the effluent(∼53.3%)after WWTP,where the Escherichia coli was removed the most,while Klebsiella pneumoniae was the most persistent.The secondary tank removed the most of FCB(by 3∼4 logs)compared to other processes,but increased all the concerned antibiotic resistant rate.The potential super bugs of FCB community showing resistance to all the target antibiotics were selected in the biological treatment unit of WWTP.The FCB showed the highest multiple antibiotic resistance(92.9%)in total which even increased to 100%in the effluent.Klebsiella has the highest antibiotic resistant rate in FCB,with a multiple antibiotic resistance rate of 98.4%.These indicated that the Klebsiella pneumoniae not just Escherichia coli should be specially emphasized after WWTP concerning the health risk associated with FCB community.

Regulatory mechanisms of dopamine metabolism in a marine Meyerozyma guilliermondii GXDK6 under NaCl stress as revealed by integrative multi-omics analysis

Dopamine can be used to treat depression,myocardial infarction,and other diseases.However,few reports are available on the de novo microbial synthesis of dopamine from low-cost substrate.In this study,integrated omics technology was used to explore the dopamine metabolism of a novel marine multi-stress-tolerant aromatic yeast Meyerozyma guilliermondii GXDK6.GXDK6 was found to have the ability to biosynthesize dopamine when using glucose as the substrate.14 key genes for the biosynthesis of dopamine were identified by whole genome-wide analysis.Transcriptomic and proteomic data showed that the expression levels of gene AAT2 encoding aspartate aminotransferase(regulating dopamine anabolism)were upregulated,while gene AO-I encoding copper amine oxidase(involved in dopamine catabolism)were downregulated under 10%NaCl stress compared with non-NaCl stress,thereby contributing to biosynthesis of dopamine.Further,the amount of dopamine under 10%NaCl stress was 2.51-fold higher than that of zero NaCl,which was consistent with the multi-omics results.Real-time fluorescence quantitative PCR(RT-qPCR)and high-performance liquid chromatography(HPLC)results confirmed the metabolic model of dopamine.Furthermore,by overexpressing AAT2,AST enzyme activity was increased by 24.89%,the expression of genes related to dopamine metabolism was enhanced,and dopamine production was increased by 56.36%in recombinant GXDK6AAT2.In conclusion,Meyerozyma guilliermondii GXDK6 could utilize low-cost carbon source to synthesize dopamine,and NaCl stress promoted the biosynthesis of dopamine.

Optimization management of hybrid energy source of fuel cell truck based on model predictive control using traffic light information

Energy optimization management can make fuel cell truck(FCT)power system more efficient,so as to improve vehicle fuel economy.When the structure of power source system and the torque distribution strategy are determined,the essence is to find the reasonable distribution of electric power between the fuel cell and other energy sources.The paper simulates the assistance of the intelligent transport system(ITS)and carries out the eco-velocity planning using the traffic signal light.On this basis,in order to further improve the energy efficiency of FCT,a model predictive control(MPC)-based energy source optimization management strategy is innovatively developed,which uses Dijkstra algorithm to achieve the minimization of equivalent hydrogen consumption.Under the scenarios of signalized intersections,based on the planned eco-velocity,the off-line simulation results show that the proposed MPC-based energy source management strategy(ESMS)can reduce hydrogen consumption of fuel cell up to 7%compared with the existing rule-based ESMS.Finally,the Hardware-in-the-Loop(HiL)simulation test is carried out to verify the effectiveness and real-time performance of the proposed MPC-based energy source optimization management strategy for the FCT based on eco-velocity planning with the assistance of traffic light information.