Development of an Antioxidant-Rich Sugar-Free Plantain Candy and Assessment of Its Shelf Life in a Flexible Laminate.

Research background: Candy is a popular confection worldwide, and it would be beneficial to society if it were converted into a source of antioxidant molecules to eliminate its adverse health effects. The amount of antioxidants available even in fruit candies is questionable due to the high thermal processing losses they undergo and the presence of various food additives. Plantains (Musa paradisiaca) are less known as good sources of biotherapeutic antioxidants, namely l-tryptophan, serotonin and melatonin, and consumption of this highly nutritious fruit is limited to underdeveloped and developing countries. The objectives of this study are: to develop a functional antioxidant-rich sugar-free plantain-based candy with valuable contents of the mentioned biomolecules in synergy; and to ensure its extended shelf life without compromising its physicochemical properties and functionality by wrapping it with a suitable packaging laminate. Experimental approach: To accomplish the first objective, lyophilized plantain powder, sorbitol and mannitol were used as base materials with minimal additives under minimal processing conditions to reduce processing loss. Sensory, proximate, physicochemical and phytochemical properties, including the antioxidant synergy among the mentioned biomolecules of the developed candies were evaluated. For the second objective, the candies were enclosed in two different flexible packaging laminates and the optimal packaging was determined based on the microbiological safety and sensory appeal of the wrapped candies. Subsequently, the above-mentioned properties of the packaged (in the most suitable laminate) candies were evaluated at regular time intervals during storage for assessment of their shelf life. Results and conclusions: The candy had a characteristic flavour of plantain, uniform dark brown colour, rich mouthfeel, pleasant aroma, moderately hard texture and moderate sweetness, along with high antioxidant activity and considerable content of l-tryptophan, serotonin and melatonin (present as a synergistic consortium). During storage of the packaged candy under ambient conditions, it remained microbiologically safe for up to 56 days, and also maintained sensory attributes, antioxidant activity and synergy compared to the control candy. Novelty and scientific contribution: This newly developed semi-hard sugar-free candy with high antioxidant content, containing three important antioxidants, namely l-tryptophan, serotonin and melatonin, could be a good source of biotherapeutic molecules and a substitute for commercial candies consumed globally.

Air pollution mitigation and suspended particulate matter retention potential of selected plant species across seasonal variation in the urban area.

Air pollution is a pressing environmental concern in urban areas, especially in densely populated cities like Delhi, India. However, plant species can effectively capture airborne suspended pollutants. Given this, the present study aimed to investigate the seasonal variations (pre- and post-monsoon) in the pollution-mitigating potential, biochemical characteristics, and suspended particulate matter (SPM) capturing capacities of select plant species in Delhi. Also, using biochemical parameters, plant morphology, and socioeconomic factors, the study computed tolerance indices such as the Air Pollution Tolerance Index (APTI) and Anticipated Performance Index (API). Ficus religiosa L. exhibited the highest APTI value of 11.94, while Polyalthia longifolia (Sonn.) Thwaites displayed the lowest 7.99 APTI value during the pre-monsoon. Ficus benghalensis L. showed the maximum SPM adhesion on the leaves, with a deposition of 1305.46 µg/cm2, whereas F. religiosa exhibited the lowest SPM deposition of 56.62 µg/cm2. Moreover, the statistical analysis indicated a positive correlation between ascorbic acid and chlorophyll content (R2 > 0.6) with APTI. Also, F. religiosa demonstrated a significant Pearson's correlation (P < 0.05) between chlorophyll content and SPM deposition during the pre-monsoon. The study highlighted the dynamic nature of plant-based air pollution mitigation. It offered valuable insights into the potential of green infrastructure as a sustainable solution for addressing air quality concerns in urban environments. The results emphasized the significance of selecting adequate plant species and considering seasonal variations in developing urban greening strategies to combat air pollution.

Effects of the Salt Stress Duration and Intensity on Developmental and Physiological Features of the Moss Polytrichum formosum.

The two accessions of the polytrichaceous moss species Polytrichum formosum, namely German and Serbian genotypes, were subjected to salt stress, aiming to study the species' developmental and physiological features. Various concentrations of sodium chloride were applied to an axenic in vitro culture of the two moss genotypes, and the growth parameters as well as physiological feature changes were followed. As inferred by the morpho-developmental parameters and survival index, the Serbian genotype showed higher resistance to salt stress as compared to the German one. However, both moss genotypes survived the highest applied concentration (500 mM). As expected, short exposures to salt were rather easily overcome. No clear patterns in sugar content and changes were observed during the stress, but they are surely included in salt stress response and tolerance in P. formosum. Longer stress increased total chlorophyll content in both genotypes. In short-term applied salt stress, the Serbian genotype had a higher total chlorophyll concentration to control unstressed plants, while the German genotype decreased the total amount of chlorophyll. Similarly, carotenoids were shown to be significantly higher in the Serbian genotype, both in unstressed and treated plants, compared to the German one. The contents of tocopherols were higher in the Serbian genotype in controlled unstressed and subsequently short- and long-stressed plantlets compared to the German accession. In general, we can assume that P. formosum is unexpectedly tolerant to salt stress and that there are differences within various accessions of overall European populations, as referred by two randomly selected genotypes, which is most probably a consequence of different genetic structure.

Comparative physiological, biochemical and transcriptomic analyses to reveal potential regulatory mechanisms in response to starvation stress in Cipangopaludina chinensis.

Cipangopaludina chinensis, as a financially significant species in China, represents a gastropod in nature which frequently encounters starvation stress owing to its limited prey options. However, the underlying response mechanisms to combat starvation have not been investigated in depth. We collected C. chinensis under several times of starvation stress (0, 7, 30, and 60 days) for nutrient, biochemical characteristics and transcriptome analyses. The results showed that prolonged starvation stress (> 30 days) caused obvious fluctuations in the nutrient composition of snails, with dramatic reductions in body weight, survival and digestive enzyme activity (amylase, protease, and lipase), and markedly enhanced the antioxidant enzyme activities of the snails. Comparative transcriptome analyses revealed 3538 differentially expressed genes (DEGs), which were significantly associated with specific starvation stress-responsive pathways, including oxidative phosphorylation and alanine, aspartate, and glutamate metabolism. Then, we identified 40 candidate genes (e.g., HACD2, Cp1, CYP1A2, and GPX1) response to starvation stress through STEM and WGCNA analyses. RT-qPCR verified the accuracy and reliability of the high-throughput sequencing results. This study provides insights into snail overwintering survival and the potential regulatory mechanisms of snail adaptation to starvation stress.

Abnormal Serum Biochemical Results and Mitochondrial Damage of Lymphocytes in Patients with Schizophrenia and SARS-CoV-2 Infection: A Retrospective Study.

Purpose: In this study, we investigated the differences in clinical biochemical values and mitochondrial mass between schizophrenia patients with and without COVID-19, so as to provide assistance to the treatment and management of COVID-19 positive patients with schizophrenia. Patients and methods: We undertook an exploratory, retrospective review of patient data from Dec. 6, 2022, to Jan. 31, 2023. A total of 1696 inpatients with psychosis (921 schizophrenia patients and 775 diagnosed with other mental diseases) during this period were identified. Finally, 60 schizophrenia patients were enrolled in our study, and 20 of them were infected with syndrome coronavirus 2 (SARS-CoV-2). The serum biochemical levels and single-cell mitochondrial mass (SCMM) of the T lymphocytes of all schizophrenia patients were analyzed. Results: The serum levels of aspartate aminotransferase (AST), alkaline phosphatase (ALP), creatinine (Cr) and lactate dehydrogenase (LDH) were significantly higher in schizophrenia patients with COVID-19 (SCZ-C) group. In addition, the SCZ-C group showed lower CD3+, CD3+CD4+ and CD3+CD8+ cell counts and higher SCMM of T lymphocytes compared to SCZ group. Furthermore, positive correlations were found between the T-cell subpopulation counts and positive symptom scores on the Positive and Negative Syndrome Scale (PANSS). Conclusion: Our study findings showed that schizophrenia patients with COVID-19 have a phenotype of mitochondrial damage in T lymphocytes and higher serum levels of AST, ALP, Cr and LDH, which might provide evidence for treating individuals with schizophrenia during subsequent spread of infectious disease.

Biochemical characterization and cleavage specificities analyses of three endo-1,3-fucanases within glycoside hydrolase family 174.

Sulfated fucans have garnered extensive research interest in recent decades due to their varied bioactivity. Fucanases are important tools for investigating sulfated fucans. This study reported the bioinformatic analysis and biochemical properties of three GH174 family endo-1,3-fucanases. Wherein, Fun174Rm and Fun174Sb showed the highest optimal reaction temperature among the reported fucanases, and Fun174Sb possessed favorable thermostability and catalysis efficiency. Fun174Rm displayed a random endo-acting manner, while Fun174Ri and Fun174Sb hydrolyzed sulfated fucan in processive manners. UPLC-MS and NMR analyses confirmed that the three enzymes catalyze cleavage of the α(1 â†’ 3)-bonds between Fucp2S and Fucp2S in the sulfated fucan from Isostichopus badionotus. These enzymes demonstrated novel cleavage specificities, which could accept α-Fucp2S residues at subsites -1 and + 1. The acquiring of these biotechnological tools would be beneficial to the in-depth research of sulfated fucans.

Morphological and biochemical characteristics associated with autophagy in gastrointestinal diseases.

Autophagy is a cellular catabolic process characterized by the formation of double-membrane autophagosomes. Transmission electron microscopy is the most rigorous method to clearly visualize autophagic engulfment and degradation. A large number of studies have shown that autophagy is closely related to the digestion, secretion, and regeneration of gastrointestinal (GI) cells. However, the role of autophagy in GI diseases remains controversial. This article focuses on the morphological and biochemical characteristics of autophagy in GI diseases, in order to provide new ideas for their diagnosis and treatment.

Physiological and biochemical effects of polystyrene micro/nano plastics on Arabidopsis thaliana.

Microplastics have garnered global attention due to their potential ecological risks. Research shows micro/nano-plastics pollution has adverse effects on plant growth, development, and physiological characteristics. However, the mechanisms underlying these effects remain unclear. The study examined the effects of polystyrene micro/nano-plastics with varying sizes and concentrations on different physiological and biochemical markers of A. thaliana. The indicators assessed include seed viability, growth, chlorophyll content, accumulation of root reactive oxygen species, and root exudates. Using fluorescence labeling, we investigated the absorption and translocation processes of micro/nano-plastics in A. thaliana. We also performed transcriptomic analysis to better understand the particular mechanisms of micro/nano-plastics. It indicated that micro/nano-plastics had an adverse effect on seed germination, especially under high concentration and small particle size treatments. This effect diminished with prolonged exposure. High concentrations at 50 nm and 100 nm treatment groups significantly inhibited the growth. Conversely, low concentrations of 1000 nm had a promoting effect. Exposure to micro/nano-plastics potentially resulted in decreased chlorophyll content, the accumulation of H2O2 in roots, and stimulated root secretion of oxalic acid. Through transcriptomic analysis, the gene expression linked to micro/nano-plastic treatments of varying sizes enriched multiple metabolic pathways, impacting plant growth, development, environmental adaptation, metabolism, pigment synthesis, and stress response.

Heterologous Expression and Characterization of a Novel Mesophilic Maltogenic α-Amylase AmyFlA from Flavobacterium sp. NAU1659.

A novel amylase AmyFlA from Flavobacterium sp. NAU1659, AmyFlA, was cloned and expressed in Esherichia coli. Based on phylogenetic and functional analysis, it was identified as a novel member of the subfamily GH13_46, sharing high sequence identity. The protein was predicted to consist of 620 amino acids, with a putative signal peptide of 25 amino acids. The enzyme was able to hydrolyze soluble starch with a specific activity of 352.97 U/mg at 50 °C in 50 mM phosphate buffer (pH 6.0). The Km and Vmax values of AmyFlA were respectively 3.15 mg/ml and 566.36 µmol·ml-1·min-1 under optimal conditions. Its activity towards starch was enhanced by 63% in the presence of 1 mM Ca2+, indicating that AmyFlA was a Ca2+-dependent amylase. Compared to the reported maltogenic amylases, AmyFlA produced a lower variety of intermediate oligosaccharides at the start of the reaction so that the product mixture contained a higher proportion of maltose. These results indicate that AmyFlA may be potential application value in the production of high-maltose syrup.

Growth, haemato-biochemical, hormonal and disease characteristics in Black Bengal goats: a review.

The goats have been considered one of the noteworthy animals to provide food security and could promote socio-economic upliftment under challenging climatic scenarios in the coming decades, particularly in the tropics. Black Bengal goat (BBG) is one of the recognised native meat-type breeds of hot-humid tropics with distinguished characteristics, including superior-quality meat, excellent skin and high prolificacy. Smaller body mass, lower metabolic rate and efficient utilisation of high-fibre forages enable BBG to adapt to a wide range of harsh climates in the tropics. The BBG can maintain physiological homeostasis efficiently in terms of electrolyte profile, endocrine functions and haemato-biochemical traits in different life phases, including the gestation period, even in high-saline coastal areas of hot-humid tropics. Crossbreeding to improve its growth rate has been attempted, but the prolificacy has been decayed. This review is intended to attract global attention to the adaptive potentialities of Black Bengal goats in terms of growth and production, haemato-biochemical, endocrinological, salt tolerance and disease characteristics that could be an asset of climate-resilient agricultural farming.

Can deep fertilization in spring maize fields improve soil properties and their distribution in soil profile?

Deep fertilization strategy has been proven to be an important fertilizer management method for improving fertilizer utilization efficiency and crop yield. However, the relationship between soil chemical and biochemical characteristics and crop productivity under different fertilization depth patterns still needs comprehensive evaluation. Field tests on spring maize were therefore carried out in the Loess Plateau of China for two successive growing seasons from 2019 to 2020. Four distinct fertilization depths of 5 cm, 15 cm, 25 cm, and 35 cm were used to systematically investigate the effects of fertilization depth on soil physicochemical parameters, enzyme activity, and biochemical properties. The findings demonstrated that although adjusting fertilization depths (D15, D25) did not significantly affect the soil organic carbon content, they did significantly improve the soil chemical and biochemical characteristics in the root zone (10-30 cm), with D25 having a greater influence than D15. Compared with D5, the total nitrogen (TN), total phosphorus (TP), available nitrogen (AN), Olsen-P, dissolved organic carbon, and nitrogen (DOC and DON) in the root zone of D25 significantly increased by 12.02%, 7.83%, 22.21%, 9.56%, 22.29%, and 26.26%, respectively. Similarly, the urease, invertase, phosphatase, and catalase in the root zone of D25 significantly increased by 9.56%, 13.20%, 11.52%, and 18.05%, while microbial biomass carbon, nitrogen, and phosphorus (MBC, MBN, and MBP) significantly increased by 18.91%, 32.01% and 26.50%, respectively, compared to D5. By optimizing the depth of fertilization, the distribution ratio of Ca2-P and Ca8-P in the inorganic phosphorus components of the root zone can also be increased. Therefore, optimizing fertilization depth helps to improve soil chemical and biochemical characteristics and increase crop yield. The results of this study will deepen our understanding of how fertilization depth influence soil properties and crop responses.

Characterization and Pathogenicity of Flavobacterium psychrophilum Isolated from Rainbow Trout (Oncorhynchus mykiss) in Korea.

Flavobacterium psychrophilum is the causative agent of bacterial cold-water disease in salmonids and rainbow trout fry syndrome. This pathogen has attained a global presence and can spread both horizontally and vertically. However, it was not documented in Korea before September 2018. In this study, the objectives were to characterize Flavobacterium psychrophilum strain FPRT1, isolated from diseased rainbow trout genotypically and phenotypically. We also conducted various investigations to better understand its impact and assess potential control measures. We acquired fifty rainbow trout (approximately 70 g in weight) and transferred them to a laboratory aquarium. During the initial acclimation period, we observed mortality and examined affected fish for clinical signs. We isolated the bacterium from the spleen of infected rainbow trout using tryptone yeast extract salts agar supplemented with glucose, naming this FPRT1. Antibiotic susceptibility testing was carried out, and from the result, we selected enrofloxacin to administer to the trout orally to reduce mortality. To evaluate pathogenicity, we exposed the trout to FPRT1 at different water temperatures (8, 15, and 22 °C). Genomic analysis was conducted to identify the serotype and relatedness of FPRT1 to European strains. Affected fish displayed clinical signs, such as ulcerative lesions in the mandible, anemia with pale gills, exophthalmia, and increased mucus secretion. Internal symptoms included pale liver and enlarged spleen. FPRT1 was susceptible to erythromycin, enrofloxacin, florfenicol, oxytetracycline, and gentamicin, but resistant to oxolinic acid and sulfamethoxazole/trimethoprim. Oral administration of enrofloxacin resulted in a decrease in mortality from 28% to 6%. Pathogenicity tests revealed varying mortality rates due to FPRT1 at different temperatures. The highest rates were observed at 8 °C (ranging from 43% to 100%) for both intraperitoneal and intramuscular injections, and lower rates occurred at 22 °C (ranging from 0% to 30%), with intramuscular injections displaying higher susceptibility. Genomic analysis identified FPRT1 as serotype 2 and indicated its close genetic relationship with European strains based on the core genome and dispensable genome. The substantial genomic similarity between our strain and European strains suggests the possibility of bacterial spread through the importation of fertilized eggs from Europe. In conclusion, this study highlights the introduction of the previously undocumented pathogen (F. psychrophilum) into Korean rainbow trout populations. The detection of this pathogen and its pathogenicity assessment is not only important for understanding its impact on local aquaculture but also for establishing surveillance and control measures to prevent further transmission and outbreaks in the region.

Clinical Characterization of Alagille Syndrome in Patients with Cholestatic Liver Disease.

Alagille syndrome (ALGS) is a multisystem condition characterized by cholestasis and bile duct paucity on liver biopsy and variable involvement of the heart, skeleton, eyes, kidneys, and face and caused by pathogenic variants in the JAG1 or NOTCH2 gene. The variable expressivity of the clinical phenotype and the lack of genotype-phenotype correlations lead to significant diagnostic difficulties. Here we present an analysis of 18 patients with cholestasis who were diagnosed with ALGS. We used an NGS panel targeting coding exons of 52 genes, including the JAG1 and NOTCH2 genes. Sanger sequencing was used to verify the mutation in the affected individuals and family members. The specific facial phenotype was seen in 16/18 (88.9%). Heart defects were seen in 8/18 (44.4%) patients (pulmonary stenosis in 7/8). Butterfly vertebrae were seen in 5/14 (35.7%) patients. Renal involvement was detected in 2/18 (11.1%) cases-one patient had renal cysts, and one had obstructive hydronephrosis. An ophthalmology examination was performed on 12 children, and only one had posterior embryotoxon (8.3%). A percutaneous liver biopsy was performed in nine cases. Bile duct paucity was detected in six/nine cases (66.7%). Two patients required liver transplantation because of cirrhosis. We identified nine novel variants in the JAG1 gene-eight frameshift variants (c.1619_1622dupGCTA (p.Tyr541X), c.1160delG (p.Gly387fs), c.964dupT (p.C322fs), c.120delG (p.L40fs), c.1984dupG (p.Ala662Glyfs), c.3168_3169delAG (p.R1056Sfs*51), c.2688delG (p.896CysfsTer49), c.164dupG (p.Cys55fs)) and one missense variant, c.2806T > G (p.Cys936Gly). None of the patients presented with NOTCH2 variants. In accordance with the classical criteria, only six patients could meet the diagnostic criteria in our cohort without genetic analysis. Genetic testing is important in the diagnosis of ALGS and can help differentiate it from other types of cholestasis.

Identification and characterization of ι-carrageenase from macroalgae-associated bacterium Microbulbifer sp. YNDZ01.

BACKGROUND: In the present study, the ι-carrageenase gene, Car1293, was obtained from the genome of Microbulbifer sp. YNDZ01, which was isolated from the surface of macroalgae. To date, there are few studies on ι-carrageenase and the anti-inflammatory activity of ι-carrageenan oligosaccharides (CGOS). To enhance our perspective on ι-carrageenase and ι-carrageen oligosaccharides, the sequence, protein structure, enzymatic properties, enzymatic digestion products and anti-inflammatory activity of the gene were investigated. RESULTS: The gene length of Car1293 is 2,589 bp, encoding an enzyme with 862 amino acids, which shares 34% similarity with any previously reported ι-carrageenase. The spatial structure of Car1293 consists of many α-helices with a ß-fold binding module located at its terminus, and eight binding sites were found in the binding module as a result of docking with CGOS-DP4 ligand. The optimum temperature and pH for the activity of recombinant Car1293 toward ι-carrageenan were 50 °C and 6.0, respectively. The hydrolysates of Car1293 are mainly degree of polymerization (DP)8, with minor products showing DP2, DP4, and DP6. The enzymatic hydrolysates CGOS-DP8 showed prominent anti-inflammatory activity, which was greater than that of the positive control l-monomethylarginine in lipopolysaccharide-induced RAW264.7 macrophages. It inhibited nitric oxide production, as well as significantly inhibited tumor necrosis factor-α and interleukin-6 secretion. CONCLUSION: The ι-carrageenase sequence encoded by Car1293 is novel and can hydrolyze carrageenan into CGOS-DP8 that has a significant anti-inflammatory effect. The present study fills a gap in the research on the biological activity of oligosaccharides in ι-carrageenan and provides promising data for the development of natural anti-inflammatory agent. © 2023 Society of Chemical Industry.

Data on cloning, expression and biochemical characteristics of a chondroitin sulfate/dermatan sulfate 4-O-endosulfatase.

The data shown in this article are related to the published paper entitled "A novel 4-O-endosulfatase with high potential for the structure-function studies of chondroitin sulfate/dermatan sulfate" in Carbohydrate Polymers. In this article, the phylogenetic analysis, cloning, expression, purification, specificity and biochemical characteristics of the identified chondroitin sulfate/dermatan sulfate 4-O-endosulfatase (endoBI4SF) are described in detail. The recombinant endoBI4SF with a molecular mass of 59.13 kDa can can specifically hydrolyze the 4-O- but not 2-O- and 6-O-sulfate groups in the oligo-/polysaccharides of chondroitin sulfate/dermatan sulfate and show the maximum reaction rate in 50 mM Tris-HCl buffer (pH 7.0) at 50°C, which can be a very useful tool for the structural and functional studies of chondroitin sulfate/dermatan sulfate.

Characteristics of antimicrobial peptide OaBac5mini and its bactericidal mechanism against Escherichia coli.

Introduction: Antimicrobial peptides (AMPs) play an important role in defending against the attack of pathogenic microorganisms. Among them, the proline-rich antibacterial peptides (PrAMPs) have been attracting close attention due to their simple structure, strong antibacterial activity, and low cell toxicity. OaBac5mini is an active fragment of the sheep-derived OaBac5 belonging to the PrAMPs family. Methods: In this study, the antibacterial activity of OaBac5mini was investigated by testing the MICs against different stains of E. coli and S. aureus as well as the time-kill curve. The bactericidal mechanism was explored by determining the effect of OaBac5mini on the cell membrane. The stability and biosafety were also evaluated. Results: The susceptibility test demonstrated that OaBac5mini showed potent antibacterial activity against the multidrug-resistant (MDR) E. coli isolates. It is noticeable that the absence of inner membrane protein SbmA in E. coli ATCC 25922 caused the MIC of OaBac5mini to increase 4-fold, implying OaBac5mini can enter into the cytoplasm via SbmA and plays its antibacterial activity. Moreover, the antibacterial activity of OaBac5mini against E. coli ATCC 25922 was not remarkably affected by the serum salts except for CaCl2 at a physiological concentration, pH, temperature, repeated freeze-thawing and proteases (trypsin < 20 µg/mL, pepsin or proteinase K). Time-kill curve analysis showed OaBac5mini at the concentration of 200 µg/mL (8 × MICs) could effectively kill E. coli ATCC 25922 after co-incubation for 12 h. In addition, OaBac5mini was not hemolytic against rabbit red blood cells and also was not cytotoxic to porcine small intestinal epithelial cells (IPEC-J2). Bioinformatic analysis indicated that OaBac5mini is a linear peptide with 8 net positive charges. Furthermore, OaBac5mini significantly increased the outer membrane permeability and impaired the inner membrane integrity and ultrastructure of E. coli ATCC25922. Conclusion: OaBac5mini is a stable and potent PrAMP that kills E. coli by two different modes of action - inhibiting intracellular target(s) and damaging cell membrane.

Cauliflower-shaped Pleurotus ostreatus cultivated in an atmosphere with high environmental carbon dioxide concentration.

Commercial aspects, physiological properties, and nutritional characteristics of Pleurotus ostreatus grown under various environmental carbon dioxide concentration ([CO2]e) conditions were assessed. As [CO2]e increased, the activity of antioxidant enzymes (catalase, peroxidase, and superoxide dismutase) in fruiting body increased, activities of succinate dehydrogenase and cytochrome c oxidase were inhibited, and malondialdehyde and adenosine triphosphate (ATP) syntheses were reduced, leading to incomplete development of pilei and stipes, or even absence of pilei. Under high [CO2]e (≥1.00%), fruiting body of P. ostreatus was morphologically altered to assume cauliflower shape. This cultivation condition resulted in high total contents of crude protein, crude fiber, and amino acids, increased levels of umami- and sweet-tasting amino acids, and reduced levels of bitter-tasting amino acids, thus enhancing the flavor of the product. In conclusion, a novel "cauliflower-shaped" mushroom (P. ostreatus) was successfully cultivated at high (≥1.00%) environmental CO2 concentration. The product has a delicious taste and high nutritional value, is relatively easy to transport and store, and has excellent potential for commercial development.

Biochemical and functional characterization of an exonuclease from Chaetomium thermophilum.

Exonucleases are often found associated with polymerase or helicase domains in the same enzyme or can function as autonomous entities to maintain genome stability. Here, we uncovered Chaetomium thermophilum RecQ family proteins that also have exonuclease activity in addition to their main helicase function. The novel exonuclease activity is separate from the helical core domain and coexists with the latter two enzymatic activities on the same polypeptide. The CtRecQ121-366 exonuclease region performs independently as an exonuclease. We describe its catalytic mechanism and biological characteristics. We demonstrate unequivocally that CtRecQ121-366 exclusively displays exonuclease activity and that this activity has a 3'-5' polarity that can both hydrolyze ssDNA and cleave dsDNA substrates. The hydrolytic activity of majority exonuclease is driven by bimetal ions, and this appears to be the case for the CtRecQ121-366 exonuclease as well. Additionally, the maximum activity of CtRecQ121-366 was observed at pH 8.0-9.0, low salt with Mg2+. The two helices in the structure, a6 and a7, play significant roles in the execution by anticipating their shape and changing essential amino acids.

Genes, Structural, and Biochemical Characterization of Four Chlorophyllases from Solanum lycopersicum.

Recent studies have confirmed that chlorophyllase (CLH), a long-found chlorophyll (Chl) dephytylation enzyme for initiating Chl catabolism, has no function in leaf senescence-related Chl breakdown. Yet, CLH is considered to be involved in fruit degreening and responds to external and hormonal stimuli. The purpose of this work was to elucidate in detail the biochemical, structural properties, and gene expression of four CLHs from the Solanum lycopersicum genome so as to understand the roles of Solanum lycopersicum chlorophyllases (SlCLHs). SlCLH1/4 were the predominantly expressed CLH genes during leaf and fruit development/ripening stages, and SlCLH1 in mature green fruit was modulated by light. SlCLH1/2/3/4 contained a highly conserved GHSXG lipase motif and a Ser-Asp-His catalytic triad. We identified Ser159, Asp226, and His258 as the essential catalytic triad by site-directed mutagenesis in recombinant SlCLH1. Kinetic analysis of the recombinant enzymes revealed that SlCLH1 had high hydrolysis activities against Chl a, Chl b, and pheophytin a (Phein a), but preferred Chl a and Chl b over Phein a; SlCLH2/3 only showed very low activity to Chl a and Chl b, while SlCLH4 showed no Chl dephytylation activity. The recombinant SlCLH1/2/3 had different pH stability and temperature optimum. Removal of the predicted N-terminal processing peptide caused a partial loss of activity in recombinant SlCLH1/2 but did not compromise SlCLH3 activity. These different characteristics among SlCLHs imply that they may have different physiological functions in tomato.

Identification of Genes and Metabolic Pathways Involved in Resin Yield in Masson Pine by Integrative Analysis of Transcriptome, Proteome and Biochemical Characteristics.

Masson pine (Pinus massoniana L.) is one of the most important resin-producing tree species in southern China. However, the molecular regulatory mechanisms of resin yield are still unclear in masson pine. In this study, an integrated analysis of transcriptome, proteome, and biochemical characteristics from needles of masson pine with the high and common resin yield was investigated. The results showed that chlorophyll a (Chl a), chlorophyll b (Chl b), total chlorophyll (Chl C), carotenoids (Car), glucose (Glu), gibberellin A9 (GA9), gibberellin A15 (GA15), and gibberellin A53 (GA53) were significantly increased, whereas fructose (Fru), jasmonic acid (JA), jasmonoyl-L-isoleucine (JA-ILE), gibberellin A1 (GA1), gibberellin A3 (GA3), gibberellin A19 (GA19), and gibberellin A24 (GA24) were significantly decreased in the high resin yield in comparison with those in the common one. The integrated analysis of transcriptome and proteome showed that chlorophyll synthase (chlG), hexokinase (HXK), sucrose synthase (SUS), phosphoglycerate kinase (PGK), dihydrolipoamide dehydrogenase (PDH), dihydrolipoamide succinyltransferase (DLST), 12-oxophytodienoic acid reductase (OPR), and jasmonate O-methyltransferases (JMT) were consistent at the transcriptomic, proteomic, and biochemical levels. The pathways of carbohydrate metabolism, terpenoid biosynthesis, photosynthesis, and hormone biosynthesis may play crucial roles in the regulation of resin yield, and some key genes involved in these pathways may be candidates that influence the resin yield. These results provide insights into the molecular regulatory mechanisms of resin yield and also provide candidate genes that can be applied for the molecular-assisted selection and breeding of high resin-yielding masson pine.