Hydrolase Activity Research Articles

Gut microbial change after administration of Lacticaseibacillus paracasei AO356 is associated with anti-obesity in a mouse model.

The status of an impaired gut microbial community, known as dysbiosis, is associated with metabolic diseases such as obesity and insulin resistance. The use of probiotics has been considered an effective approach for the treatment and prevention of obesity and related gut microbial dysbiosis. The anti-obesity effect of Lacticaseibacillus paracasei AO356 was recently reported. However, the effect of L. paracasei AO356 on the gut microbiota has not yet been identified. This study aimed to elucidate the effect of L. paracasei AO356 on gut microbiota and ensure its safety for use as a probiotic. Oral administration of L. paracasei AO356 (107 colony-forming units [CFU]/mg per day, 5 days a week, for 10 weeks) to mice fed a high-fat diet significantly suppressed weight gain and fat mass. We investigated the composition of gut microbiota and explored its association with obesity-related markers. Oral administration of L. paracasei AO356 significantly changed the gut microbiota and modified the relative abundance of Lactobacillus, Bacteroides, and Oscillospira. Bacteroides and Oscillospira were significantly related to the lipid metabolism pathway and obesity-related markers. We also confirmed the safety of L. paracasei AO356 using antibiotics resistance, hemolysis activity, bile salt hydrolase activity, lactate production, and toxicity tests following the safety assessment guidelines of the Ministry of Food and Drug Safety (MFDS). This study demonstrated that L. paracasei AO356 is not only associated with an anti-obesity effect but also with changes in the gut microbiota and metabolic pathways related to obesity. Furthermore, the overall safety assessment seen in this study could increase the potential use of new probiotic materials with anti-obesity effects.

Induction and Characterisation of Lignocellulolytic Activities from Novel Deep-Sea Fungal Secretomes

Fungi are increasingly recognised as being able to inhabit extreme environments. The deep sea is considered an extreme environment because of its low temperatures, high hydrostatic and lithostatic pressures, 3.5% salinity, and low oxygen, nutrient and light availability. Fungi inhabiting the deep sea may have evolved to produce proteins that allow them to survive these conditions. Investigation and characterisation of fungal lignocellulolytic enzymes from extreme environments like the deep sea is needed, as they may have unusual adaptations that would be useful in industry. This work, therefore, aimed to profile in detail the lignocellulolytic capabilities of fungi isolated from deep-sea sediments in the Atlantic Ocean, and a comparative lignocellulolytic terrestrial isolate. The isolates were strains of Emericellopsis maritima, Penicillium chrysogenum, P. antarcticum and Talaromyces stollii. Lignocellulolytic enzyme induction was achieved using liquid-state fermentation (LSF) with wheat bran as the main carbon source, while enzyme characteristics were evaluated using biochemical assays and gel-based proteomics. This study revealed that the isolates were halotolerant, produced xylanase over wide pH and temperature ranges, and produced a variety of glycoside hydrolase and feruloyl esterase activities. The T. stollii secretome demonstrated remarkable levels of exo-glycoside hydrolase activity, with xylanase activity optimum between pH 1.5–6.0 and temperatures between 1–60 °C, making this isolate an ideal candidate for biotechnological applications. This study is the first to quantitatively characterise xylanase activities and exo-glycoside hydrolase activities secreted by E. maritima, P. antarcticum and a marine T. stollii strain. This study is also the first to quantitatively characterise xylanase activities by a marine strain of P. chrysogenum during LSF.

Specnuezhenide Ameliorates Age-Related Hepatic Lipid Accumulation via Modulating Bile Acid Homeostasis and Gut Microbiota in D-Galactose-Induced Mice.

Age-related hepatic lipid accumulation has become a major health problem in the elderly population. Specnuezhenide (SPN) is a major active iridoid glycoside from an edible herb Fructus Ligustri Lucidi, which is commonly used for preventing age-related diseases. However, the beneficial effects of SPN on age-related liver injury remain unknown. This study aimed to reveal the effect of SPN on age-related hepatic lipid accumulation and the underlying mechanism. D-galactose (D-gal)-induced aging mice were treated with vehicle or SPN for 12 weeks. Treatment of SPN decreased lipid accumulation and inflammation in the liver of D-gal-induced mice. Untargeted and targeted metabolomics showed that the SPN could regulate the bile acid (BA) synthesis pathway and restore the BA compositions in serum, livers, and feces of the D-gal-induced mice. Furthermore, SPN enhanced the protein and mRNA levels of hepatic BAs synthesis enzymes cytochrome P45027A1, cytochrome P4507A1, cytochrome P4507B1, and cytochrome P4508B1. Meanwhile, SPN alleviated D-gal-induced gut dysbiosis and reversed the proportions of microbes associated with bile salt hydrolase activity, including Lactobacillus, Ruminiclostridium, and Butyrivibrio. Our study revealed that SPN attenuated age-related hepatic lipid accumulation by improving BA profiles via modulating hepatic BA synthesis enzymes and gut microbiota.

Identification and distribution of new candidate T6SS effectors encoded in Salmonella Pathogenicity Island 6.

The type VI secretion system (T6SS) is a contact-dependent contractile multiprotein apparatus widely distributed in Gram-negative bacteria. These systems can deliver different effector proteins into target bacterial and/or eukaryotic cells, contributing to the environmental fitness and virulence of many bacterial pathogens. Salmonella harbors five different T6SSs encoded in different genomic islands. The T6SS encoded in Salmonella Pathogenicity Island 6 (SPI-6) contributes to Salmonella competition with the host microbiota and its interaction with infected host cells. Despite its relevance, information regarding the total number of effector proteins encoded within SPI-6 and its distribution among different Salmonella enterica serotypes is limited. In this work, we performed bioinformatic and comparative genomics analyses of the SPI-6 T6SS gene cluster to expand our knowledge regarding the T6SS effector repertoire and the global distribution of these effectors in Salmonella. The analysis of a curated dataset of 60 Salmonella enterica genomes from the Secret6 database revealed the presence of 23 new putative T6SS effector/immunity protein (E/I) modules. These effectors were concentrated in the variable regions 1 to 3 (VR1-3) of the SPI-6 T6SS gene cluster. VR1-2 were enriched in candidate effectors with predicted peptidoglycan hydrolase activity, while VR3 was enriched in candidate effectors of the Rhs family with C-terminal extensions with predicted DNase, RNase, deaminase, or ADP-ribosyltransferase activity. A global analysis of known and candidate effector proteins in Salmonella enterica genomes from the NCBI database revealed that T6SS effector proteins are differentially distributed among Salmonella serotypes. While some effectors are present in over 200 serotypes, others are found in less than a dozen. A hierarchical clustering analysis identified Salmonella serotypes with distinct profiles of T6SS effectors and candidate effectors, highlighting the diversity of T6SS effector repertoires in Salmonella enterica. The existence of different repertoires of effector proteins suggests that different effector protein combinations may have a differential impact on the environmental fitness and pathogenic potential of these strains.

The presence of blastocyst within the uteri facilitates lumenal epithelium transformation for implantation via upregulating lysosome proteostasis activity

ABSTRACT The mammalian endometrium is covered by the lumenal epithelium (Le), which directly interacts with the blastocyst and plays an important role in the establishment of reciprocal crosstalk between the embryo and receptive uterus during implantation. However, the effect of the blastocyst on uterine differentiation during the window of receptivity is far from well understood. Through transcriptomic profiling of the uterine Le isolated by laser capture microdissection (LCM), it was demonstrated that global gene expression changes occurred in Le between pseudopregnant mice without embryos and pregnant mice with embryos. Some differentially expressed genes, including upregulated Areg (amphiregulin), Ihh (Indian hedgehog), Lifr (leukemia inhibitory factor receptor) and downregulated Msx1 (msh homeobox 1), Pgr (progesterone receptor), and Gata2 (GATA binding protein 2) in pregnant mice, have been reported to regulate the establishment of uterine receptivity. Besides, we found that blastocysts induced an increase in both the number and acidification of lysosome, consistent with enhanced lysosomal hydrolase activity in uterine Le. Further exploration uncovered that blastocyst-derived IGF2 was involved into the activation of epithelial STAT3 to induce lysosomal hydrolase expression, and inhibition of lysosomal function derails both uterine receptive maker gene expressions and embryo implantation. Finally, based on the proteomic data of both epithelia and the separated lysosome, it was revealed that CLDN1 (claudin 1) and MUC1 (mucin 1, transmembrane), two well-known downregulated molecules for successful implantation, are degraded by epithelial lysosome. In brief, our data demonstrated that blastocysts induced normal epithelium differentiation with lysosome activation to promote the uterine epithelial differentiation for embryo implantation. Abbreviations ACTB: actin beta; AREG: amphiregulin; ATP6V0A4: ATPase, H+ transporting, lysosomal V0 subunit A4; Baf A1: bafilomycin A1; BSA: bovine serum albumin; CLDN1: claudin 1; CTSB: cathepsin B; DEGs: differentially expressed genes; E2: 17β-estradiol; ESR: estrogen receptor; GATA2: GATA binding protein 2; GLA: galactosidase, alpha; GO: gene ontology; HBEGF: heparin-binding EGF-like growth factor; IGF1R: insulin-like growth factor 1 receptor; Ihh: Indian hedgehog; ISH: in situ hybridization; LAMP1: lysosomal-associated membrane protein 1; LCM: laser capture microdissection; Le: lumenal epithelium; LGMN: legumain; LIF: leukemia inhibitory factor; LIFR: LIF receptor alpha; MSX1: msh homeobox 1; MUC1: mucin 1, transmembrane; P4: progesterone; PBS: phosphate-buffered saline; PCA: principal component analysis; PPT1: palmitoyl-protein thioesterase 1; PGR: progesterone receptor; PSP: pseudopregnancy; PTGS2/COX2: prostaglandin-endoperoxide synthase 2; qPCR: quantitative real-time polymerase chain reaction; SP: pregnancy; TFEB: transcription factor EB.

GM1 Gangliosidosis Revealed by Fetoplacental Hydrops Syndrome: A Case Report

GM1 gangliosidos is describes a family of disorders that result from deficient activity of the lysosomal hydrolase, β-galactosidase, and which manifest varying degrees of neurodegeneration, retinal cherry-red spots, and visceromegaly. The three traditional subgroups of infantile, juvenile, and adult GM1 gangliosidosis show phenotypic overlap, although in general, the earlier the onset, the more severe the manifestations. This article gives an overview of the clinical signs in a patient with GM1 type 1 gangliosidosis revealed by fetoplacental hydrops syndrome.

Construction of covalent organic framework nanozymes with photo-enhanced hydrolase activities for colorimetric sensing of organophosphorus nerve agents

Construction of covalent organic frameworks (COFs)-based nanozymes is of great importance for the extensive applications in catalysis and sensing fields. In this work, a two-dimensional COF (DAFB-DCTP COF) was fabricated via Knoevenagel condensation reaction. The integration of catalytically active sites of pyridine groups into the donor-acceptor (D-A) conjugated skeleton endows DAFB-DCTP COF with both hydrolytic and photosensitive properties. The DAFB-DCTP COF can be utilized as an artificial enzyme with selective and photo-enhanced catalytic efficiency, facilitating its application in photocatalytic degradation of hydrolase substrates (p-nitrophenyl acetate, pNPA) by nucleophilic reaction and further realizing colorimetric detection of the nanozyme inhibitor of organophosphorus nerve agent (diethyl cyanophosphonate, DCNP). The distinct color changes could be distinguished by naked eyes even at a low DCNP concentration, and the versatile smartphone analysis featured with reliability and simplicity. For the first time, the COFs' intrinsic hydrolase activity depending on their structural characteristics was investigated in synergy with the photosensitive performance originating from their photoelectric features. The present contribution provides a promising direction towards construction of colorimetric sensing platform based on the regulation of COFs’ non-oxidoreductase activity under visible light irradiation.

Hepatocellular carcinoma cell differentiation trajectory predicts immunotherapy, potential therapeutic drugs, and prognosis of patients.

The aim of this study is to explore a novel classification and investigate the clinical significance of hepatocellular carcinoma (HCC) cells. We analyzed integrated single-cell RNA sequencing and bulk RNA-seq data obtained from HCC samples. Cell trajectory analysis divided HCC cells into three subgroups with different differentiation states: state 1 was closely related to phosphoric ester hydrolase activity, state 2 was involved in eukaryotic initiation factor 4E binding, translation regulator activity and ribosome, and state 3 was associated with oxidoreductase activity and metabolism. Three molecular classes based on HCC differentiation-related genes (HDRGs) from HCC samples were identified, which revealed immune checkpoint gene expression and overall survival (OS) of HCC patients. Moreover, a prognostic risk scoring (RS) model was generated based on eight HDRGs, and the results showed that the OS of the high-risk group was worse than that of the low-risk group. Further, potential therapeutic drugs were screened out based on eight prognostic RS-HDRGs. This study highlights the importance of HCC cell differentiation in immunotherapy, clinical prognosis, and potential molecular-targeted drugs for HCC patients, and proposes a direction for the development of individualized treatments for HCC.

Transcriptome Analysis Reveals Changes in Whole Gene Expression, Biological Process, and Molecular Functions Induced by Nickel in Jack Pine (Pinus banksiana).

Understanding the genetic response of plants to nickel stress is a necessary step to improving the utility of plants in environmental remediation and restoration. The main objective of this study was to generate whole genome expression profiles of P. banksiana exposed to nickel ion toxicity compared to reference genotypes. Pinus banksiana seedlings were screened in a growth chamber setting using a high concentration of 1600 mg of nickel per 1 kg of soil. RNA was extracted and sequenced using the Illumina platform, followed by de novo transcriptome assembly. Overall, 25,552 transcripts were assigned gene ontology. The biological processes in water-treated samples were analyzed, and 55% of transcripts were distributed among five categories: DNA metabolic process (19.3%), response to stress (13.3%), response to chemical stimuli (8.7%), signal transduction (7.7%) and response to biotic stimulus (6.0%). For molecular function, the highest percentages of genes were involved in nucleotide binding (27.6%), nuclease activity (27.3%) and kinase activity (10.3%). Sixty-two percent of genes were associated with cellular compartments. Of these genes, 21.7% were found in the plasma membrane, 16.1% in the cytosol, 12.4% with the chloroplast and 11.9% in the extracellular region. Nickel ions induced changes in gene expression, resulting in the emergence of differentially regulated categories. Overall, there were significant changes in gene expression with a total 4128 genes upregulated and 3754 downregulated genes detected in nickel-treated genotypes compared to water-treated control plants. For biological processes, the highest percentage of upregulated genes in plants exposed to nickel were associated with the response to stress (15%), the response to chemicals (11,1%), carbohydrate metabolic processes (7.4%) and catabolic processes (7.4%). The largest proportions of downregulated genes were associated with the biosynthetic process (21%), carbohydrate metabolic process (14.3%), response to biotic stimulus (10.7%) and response to stress (10.7%). For molecular function, genes encoding for enzyme regulatory and hydrolase activities represented the highest proportion (61%) of upregulated gene. The majority of downregulated genes were involved in the biosynthetic processes. Overall, 58% of upregulated genes were located in the extracellular region and the nucleus, while 42% of downregulated genes were localized to the plasma membrane and 33% to the extracellular region. This study represents the first report of a transcriptome from a conifer species treated with nickel.

Short synthesis of a broadly Reactive, cell permeable serine hydrolase Fluorophosphonate-Alkyne probe

An abbreviated synthesis of the cell permeable fluorophosphonate-alkyne probe (FP-alkyne) for the broad assessment of serine hydrolase activity has been developed. While FP-alkyne has proven pivotal in numerous chemical biology studies access has relied on a lengthy preparation over nine steps. We have developed a four-step synthesis, starting from commercially available compounds, with three purification steps to provide a new expedited route allowing easy access to a useful tool compound for exploring serine hydrolases chemistry and biology. This route was used in our own studies to generate FP-alkyne which in turn was used to identify the enzyme responsible for Fatty Acid Esters of Hydroxy Fatty Acids (FAHFA) biosynthesis. The use of this route can enable the syntheses of new tool compounds in addition to improving accessibility to FP-alkyne.

Composition, distribution, health risks, and drivers of phthalates in typical red paddy soils.

The accelerated accumulation of phthalate esters (PAEs) in paddy soils poses a serious threat to human health. However, related studies mainly focus on facility vegetable fields, drylands, and orchards, and little is known about paddy soils. In this study, 125 samples were collected from typical red paddy fields to investigate the pollution characteristics, sources, health risks, and main drivers of PAEs. Soil physicochemical properties, enzyme activity, and bacterial community composition were also measured simultaneously. The results showed that eight PAE congeners were detected ranging from 0.17 to 1.97 mg kg-1. Di-n-butyl phthalate (DBP), di-(2-ethylhexyl) phthalate (DEHP), and di-isobutyl phthalate (DIBP) were the most abundant PAE congeners, accounting for 81% of the total PAEs. DEHP exhibited a potential carcinogenic risk to humans through the intake route. The main PAEs were positively correlated with soil organic matter (SOM) and soil water content (SWC) contents. Low levels of PAEs increased bacterial abundance. Furthermore, most PAE congeners were positively correlated with hydrolase activity. Soil acidity and nutrient dynamics played a dominant role in the bacterial community composition, with PAE congeners playing a secondary role. These findings suggest that there may be a threshold response between PAEs and organic matter and nutrient transformation in red paddy soils, and that microbial community should be the key driver. Overall, this study deepens the understanding of ecological risks and microbial mechanisms of PAEs in red paddy soils.

Genome-wide identification and analysis of a cotton secretome reveals its role in resistance against Verticillium dahliae

BackgroundThe extracellular space between the cell wall and plasma membrane is a battlefield in plant-pathogen interactions. Within this space, the pathogen employs its secretome to attack the host in a variety of ways, including immunity manipulation. However, the role of the plant secretome is rarely studied for its role in disease resistance.ResultsHere, we examined the secretome of Verticillium wilt-resistant Gossypium hirsutum cultivar Zhongzhimian No.2 (ZZM2, encoding 95,327 predicted coding sequences) to determine its role in disease resistance against the wilt causal agent, Verticillium dahliae. Bioinformatics-driven analyses showed that the ZZM2 genome encodes 2085 secreted proteins and that these display disequilibrium in their distribution among the chromosomes. The cotton secretome displayed differences in the abundance of certain amino acid residues as compared to the remaining encoded proteins due to the localization of these putative proteins in the extracellular space. The secretome analysis revealed conservation for an allotetraploid genome, which nevertheless exhibited variation among orthologs and comparable unique genes between the two sub-genomes. Secretome annotation strongly suggested its involvement in extracellular stress responses (hydrolase activity, oxidoreductase activity, and extracellular region, etc.), thus contributing to resistance against the V. dahliae infection. Furthermore, the defense response genes (immunity marker NbHIN1, salicylic acid marker NbPR1, and jasmonic acid marker NbLOX4) were activated to varying degrees when Nicotina benthamiana leaves were agro-infiltrated with 28 randomly selected members, suggesting that the secretome plays an important role in the immunity response. Finally, gene silencing assays of 11 members from 13 selected candidates in ZZM2 displayed higher susceptibility to V. dahliae, suggesting that the secretome members confer the Verticillium wilt resistance in cotton.ConclusionsOur data demonstrate that the cotton secretome plays an important role in Verticillium wilt resistance, facilitating the development of the resistance gene markers and increasing the understanding of the mechanisms regulating disease resistance.

Advances in Synthetic Biology Techniques and Industrial Applications of Corynebacterium glutamicum

Corynebacterium glutamicum is a Gram-positive bacterium (non-spore-forming) that has been wildly used for amino acid production. Due to its stable protein secretion, low extracellular hydrolase activity, and non-toxicity, the application field of C. glutamicum has been greatly expanded. Currently, gene editing technology based on synthetic biology has great potential for synthetic biology research and genetic modification in C. glutamicum because of its ability to efficiently regulate the physiological and metabolic networks of the strain. Therefore, we summarize the gene editing tools and strategies of C. glutamicum from the aspects of genetic modification and expression elements, and we also describe the effects of gene editing techniques on a variety of products such as amino acids and their derivatives, recombinant proteins, and functional sugars, which provide a certain theoretical basis for the research on the modification of C. glutamicum strains and industrial applications. Finally, we prospect the design and industrial application of C. glutamicum genetic modification from multiple perspectives based on gene editing techniques.

Links between regional and depth patterns of microbial communities and enzyme activities in the western North Atlantic Ocean

Heterotrophic bacteria process much of the organic matter produced by phytoplankton in the ocean. A large proportion of this organic matter is in the form of polysaccharides, structurally complex, high molecular weight sugars. To consume this complex organic matter, microbes must initially produce enzymes of the correct structural specificity to transform it to smaller pieces that can be brought into the cell. The extent to which bacteria can hydrolyze organic matter determines how much carbon is transformed by a given microbial community. Because microbial community composition differs with location and depth in the ocean, quantifying their enzymatic potential at different stations and depths is essential to reveal patterns in microbial functional capabilities. To investigate links between patterns of bacterial composition and function, we assessed the bacterial community composition and measured glucosidase, peptidase, and polysaccharide hydrolase activities throughout the water column at 16 stations in the western North Atlantic. We found that bacterial community composition and polysaccharide hydrolase activities were depth stratified and showed regional variability, while glucosidase and peptidase activities were more similar among locations and depths. These findings suggest that polysaccharide hydrolase activities are expressed by a narrower range of organisms within bacterial communities, while the abilities to degrade peptides occurs more widely among community members, likely due to the broader substrate specificity for proteins compared to polysaccharides. The data and findings presented here highlight the extent to which patterns of microbial community composition and function and the physical oceanography of the western North Atlantic are interwoven and contribute to the overall transformation of carbon in the ocean.

CB1 receptor expression and signaling are required for dexamethasone-induced aversive memory consolidation

The molecular processes that underlie long-term memory formation involve signaling pathway activation by neurotransmitter release, which induces the expression of immediate early genes, such as Zif268, having a key role in memory formation. In this work, we show that the cannabinoid CB1 receptor signaling is necessary for the effects of dexamethasone on the behavioral response in an inhibitory avoidance task, on dexamethasone-induced ERK phosphorylation, and on dexamethasone-dependent Zif268 expression. Furthermore, we provide primary evidence for the mechanism responsible for this crosstalk between cannabinoid and glucocorticoid-mediated signaling pathways, showing that dexamethasone regulates endocannabinoid metabolism by inhibiting the activity of the Fatty acid amide hydrolase (FAAH), an integral membrane enzyme that hydrolyzes endocannabinoids and related amidated signaling lipids. Our results provide novel evidence regarding the role of the endocannabinoid system, and in particular of the CB1 receptor, as a mediator of the effects of glucocorticoids on the consolidation of aversive memories.

Fiber consumption stimulates the activity of microbial bile salt hydrolases

Fiber consumption stimulates the activity of microbial bile salt hydrolases

Identification of intestinal metabolic activation of loganin generated dialdehyde reactive intermediates improves intestinal bile salt hydrolase activities

Loganin is an iridoid with potent pharmacological effects. Loganin contains a hemiacetal structure and can convert to dialdehyde intermediates after deglycosylation. We hypothesized that the metabolites of loganin with hemiacetal can generate reactive dialdehyde intermediates. This study aims to characterize the metabolic profiling of loganin and especially for the unstable dialdehyde intermediates by using ultra-performance liquid chromatograph-quadrupole orbitrap mass spectrometry. In this study, a total of 26 stable metabolites were identified in loganin-treated rats. Loganin underwent different metabolism in the intestine and liver, which was confirmed mainly by the metabolites in the hepatic portal vein. In the intestine, the major metabolic pathways were ester hydrolysis and deglycosylation, followed by methylation and dehydrogenation. The hepatic metabolism pathways were hydrogenation, hydroxylation, glucuronidation, and sulfonation. The circulating metabolites with high abundance were mainly derived from intestinal metabolism. Importantly, 11 unstable dialdehyde intermediates of loganin were identified and described for the first time. The dialdehyde intermediates were identified by their dihydropyridine conjugates with amino acids. The dialdehyde intermediates were mainly produced in the intestine. The dialdehyde intermediates enable covalent modification of intestinal proteins. Loganin can up-regulate the activity of intestinal bile salt hydrolase (BSH), catalyzing bile acid metabolism. The level of protein adducts was positively associated with BSH activity, indicating dialdehyde intermediates played a key role in the up-regulation of BSH activities. In conclusion, this study not only demonstrates the characteristic metabolic fate of loganin but also facilitates the understanding of the pharmacologic effects of dialdehyde intermediates.

Response of Soil Absolute and Specific Enzyme Activities to Long-Term Application of Organic Fertilizer to Solonetz in Northeast China

Soil organic carbon (SOC) and microbial biomass carbon (MBC) are highly correlated with enzyme activities. Specific enzyme activities can exclude the autocorrelation between enzyme activity and SOC and MBC. However, the responses of absolute and specific enzyme activities to saline–alkali properties remains unclear. In this study, the absolute and specific enzyme activities of cellobiose hydrolase, β-glucosidase, arylsulfatase, alkaline phosphatase, and urease were measured in soils with 10, 15, 18, 21, and 26 years of organic fertilizer application in contrast to soils without organic fertilizer application. The results showed that long-term organic fertilizer application led to significantly increased in the absolute and specific enzyme activity and decrease in pH, electrical conductivity (EC), exchangeable sodium percentage (ESP), as well as sodium adsorption ratio (SAR5:1). In the structural equation model (SEM), the EC extremely limited the geometric mean of specific enzyme activity per unit of MBC (MBC-GMSEA) (path coefficient, −0.84, p < 0.001). Pearson’s correlation analysis showed that the correlations between EA/MBC and pH, EC, ESP, and SAR5:1 were higher than between the absolute soil enzyme activity and pH, EC, ESP, and SAR5:1. Of the parameters tested, EA/MBC was a more sensitive index to reflect the improvement effect of organic fertilizer on soils and evaluate the saline–alkali barrier.

Hydrolase Activities of Sourdough Microorganisms

Sourdough is renowned for improving bakery products’ nutritional and quality characteristics through the enzymes produced by its microbiota. Among the enzymatic framework present in sourdough fermentation, amylase, cellulase, and peptidase are responsible for many of the properties valued in sourdough products. Furthermore, there is an increasing concern regarding the allergenic potential of gluten, which motivates the investigation of enzymatic gluten hydrolysis. This study aimed to select probiotics, isolate and identify microorganisms from sourdough, and assess their amylase, cellulase, and peptidase profiles. Additionally, a rapid screening method was developed for gluten and wheat flour hydrolysis, and gluten zymography and enzymography were performed. As a result, 18 microorganisms were isolated from sourdough and identified. The probiotic Bacillus licheniformis LMG-S 28935, and three microorganisms isolated from sourdough, the Limosilactobacillus fermentum, Pediococcus pentosaceus, and Saccharomyces cerevisiae, completed the profile of analyzed hydrolases and presented the capacity to hydrolyze gluten. These findings contribute to a better understanding of sourdough microorganisms’ hydrolase activities in the bakery science and technology field. In addition, an efficient, fast, and economical method for screening extracellular glutenase, produced by microorganisms, was applied. To our knowledge, it was the first time that amylase, cellulase, and peptidase activities were assessed from sourdough microorganisms.

Potential of Y. lipolytica epoxide hydrolase for efficient production of enantiopure (R)-1,2-octanediol

The recombinant Yleh from a tropical marine yeast Yarrowia lipolytica NCIM 3589 exhibited a high epoxide hydrolase activity of 9.34 ± 1.80 µmol min-1 mg-1 protein towards 1,2-epoxyoctane (EO), at pH 8.0 and 30 °C. The reaction product was identified as 1,2-Octanediol (OD) by GC-MS using EO and H2O18 as substrate, affirming the functionality of Yleh as an epoxide hydrolase. For EO, the Km, Vmax, and kcat/Km values were 0.43 ± 0.017 mM, 0.042 ± 0.003 mM min-1, and 467.17 ± 39.43 mM-1 min-1, respectively. To optimize the reaction conditions for conversion of racemic EO by Yleh catalyst to enantiopure (R)-1,2-octanediol, initially, Response Surface Methodology was employed. Under optimized reaction conditions of 15 mM EO, 150 µg purified Yleh at 30 °C a maximal diol production of 7.11 mM was attained in a short span of 65 min with a yield of 47.4%. Green technology using deep eutectic solvents for the hydrophobic substrate (EO) were tested as co-solvents in Yleh catalyzed EO hydrolysis. Choline chloride-Glycerol, produced 9.08 mM OD with an increased OD yield of 60.5%. Thus, results showed that deep eutectic solvents could be a promising solvent for Yleh-catalyzed reactions making Yleh a potential biocatalyst for the biosynthesis of enantiopure synthons.