High Enzyme Activity Research Articles

Prognostic significance of LOXL2 enzyme activity in primary myelofibrosis.

Primary myelofibrosis (PMF) is characterized by bone marrow fibrosis, but the underlying molecular mechanisms remain incompletely understood. Here, we investigated the role of lysyl oxidase-like 2 (LOXL2), an enzyme involved in extracellular matrix remodeling, in PMF pathogenesis. Analysis of bone marrow cells from PMF patients revealed significantly elevated LOXL2 mRNA expression compared to healthy controls, which was further validated using 2 independent Gene Expression Omnibus datasets (GSE26049 and GSE12234). Serum LOXL2 protein levels were markedly increased in PMF patients (n = 38) compared to controls (n = 15) (P < .01), accompanied by significantly enhanced enzymatic activity (P < .05). Clinical analysis demonstrated that high LOXL2 enzyme activity correlated with adverse clinical features, including higher risk scores in both International Prognostic Scoring System (P = .029) and Dynamic International Prognostic Scoring System (P = .012) systems, and increased peripheral blood blast percentages (≥1%, P = .019). Notably, elevated LOXL2 enzyme activity emerged as an independent adverse prognostic factor in multivariate analysis. Gene set enrichment analysis revealed LOXL2's association with multiple biological processes, including extracellular matrix remodeling, inflammatory responses, and JAK-STAT signaling pathways. Using CellMiner drug prediction analysis, we identified several FDA-approved drugs, particularly kinase inhibitors, as potential LOXL2-targeting therapeutics. Our findings demonstrate LOXL2's critical role in PMF pathogenesis and suggest its potential as both a prognostic marker and therapeutic target in PMF treatment.

The food application of a novel Staphylococcus aureus bacteriophage vB_SA_STAP152 and its endolysin LysP152 with high enzymatic activity under cold temperature

The food application of a novel Staphylococcus aureus bacteriophage vB_SA_STAP152 and its endolysin LysP152 with high enzymatic activity under cold temperature

Biofilm-based immobilized fermentation of engineered Komagataella phaffii for xylanase production

This study presented an immobilized fermentation process of engineered Komagataella phaffii with improved biofilm-forming abilities for continuous xylanase production and provided the first insights into the molecular basis of biofilm-based immobilized fermentation of K. phaffii. Overexpression of PAS_chr2-2_0178 and PAS_FragB_0067 in K. phaffii facilitated biofilm formation with 31.6% and 113.8% increasement, respectively. Subsequently, a biofilm-based immobilized fermentation process was developed for the PAS_FragB_0067-overexpressing strain. Xylanase production over five batches by GS115-0067* was better than that of GS115-xyn, with an overall average of 35.4% higher enzyme activity. PAS_FragB_0067 overexpression resulted in better adhesion of K. phaffii cells on the carrier, and enhanced biofilms could provide more active cells in the immobilized fermentation process. Transcriptome analysis revealed that overexpression of the biofilm-related gene promoted central carbon metabolism. These findings offer a valuable reference strategy to improve production efficiency of K. phaffii cells in continuous fermentation processes.

Relevance of Neutralizing Antibodies for the Pharmacokinetics of Pegunigalsidase Alfa in Patients with Fabry Disease.

Pegunigalsidase alfa is a newly approved drug for the treatment of Fabry disease, designed to increase the plasma half-life and reduce immunogenicity of infused α-galactosidase A (AGAL). We provide the first comprehensive pharmacokinetic and immunogenic data apart from industry-initiated studies. Pharmacokinetics of pegunigalsidase alfa, amino acid, and polyethylene glycol (PEG)-specific antibodies and immunecomplexes were measured in treated patients (11 switched, two naïve). Measurements were performed in serum samples drawn directly before and after infusions over three to ten consecutive infusions. Only three patients started directly with 1.0 mg/kg body weight. No infusion-associated reactions were reported under pegunigalsidase alfa during the observation. Patients without pre-existing neutralizing anti-AGAL antibodies showed high enzymatic AGAL peak activities and sustained AGAL serum concentrations until the next infusion, which was not observed in those with neutralizing anti-AGAL antibodies. Nine (69.2%) patients presented with pre-existing anti-PEG antibodies (IgG or IgM), which seemed to have no impact on pharmacokinetics during the observation. No new anti-PEG or anti-AGAL antibody formation was observed after treatment initiation. Three (75.0%) patients with pre-existing neutralizing anti-AGAL antibodies showed a titer increase and one (25.0%) patient a decrease. In patients with anti-AGAL antibodies (n = 4) immune-complex formation was detected. The pharmacokinetics of pegunigalsidase alfa show different profiles depending on the presence of pre-existing neutralizing antibodies, with reduced plasma half-life and peak enzyme activity after infusion in patients with antibodies. The clinical significance of a reduced pegunigalsidase alfa half-life and the formation of immune complexes in antibody-positive patients needs to be analyzed in future studies.

Foraminifera and geomicrobiology as indicators of the environmental recovery in a mangrove affected by oil spills in the Guanabara Bay (Brazil)

Foraminifera could be used as indicators to evaluate the impacts of oil spills because the crude oil causes several disturbances on the development and reproduction of the species. However, little is known about the relationship between mangrove species and this pollutant. Foraminiferal assemblages were studied in 22 surface sediment samples collected from a mangrove in Guanabara Bay (Brazil) that was severely impacted by an oil spill six years earlier. The foraminiferal responses to the environmental stress helped to determine the most degraded areas. Elevated concentrations of aromatic compounds and potentially bioavailable heavy metals, with contamination levels up to nine times higher compared to the pre-industrial period in the Guanabara Bay. The dominance of anaerobic bacteria, and high esterase enzyme activity (ESTE) suggest that the bacterial community is metabolizing the hydrocarbons in the sediments. Despite these stressors, density and diversity of living foraminifera are comparable to values observed in less impacted Brazilian mangroves. Species distribution patterns reveal an environmental gradient across the mangrove with numerous species increasing their relative abundance towards the areas topographically lower and with higher levels of pollutants and ESTE. Specifically, Caronia exilis, Tiphotrocha comprimata, Ammobaculites exiguus, Textularia paranaguaensis, Ammotium cassis, Ammobaculites dilatatus, Polyssaccamina hipohalina, Siphotrochammina lobata, Ammonia tepida, Ammotium morenoi, Miliammina fusca, Entzia macrescens, Trochammina squamata and Paratrochammina clossi are inferred as pollution-tolerant taxa. This integrated geochemical, microbiological and ecological approach applied to assess the sediment quality in a complex ecosystem has important implications for the use of living foraminifera in recovery stage assessments and biomonitoring plans.

Insect-derived bacteria as biocontrol tool and a potent suppressor of plant pathogenic fungi in tomato cultivation

Sustainable agriculture is increasingly emphasized, focusing on microorganisms' role in maintaining soil fertility and inhibiting plant pathogens. Seeking novel sources of plant-beneficial bacteria, our study explores insects due to their established associations with plants and bacteria. The insect gut, hosting various bacteria, may hold microbes protecting against fungal infections, particularly plant pathogens. Traditional sources of plant growth-promoting bacteria are the rhizosphere and host plant tissues; however, insects serve as diverse bacterial reservoirs in the environment. This study aimed to identify insect-gut-derived bacteria with antifungal properties and cellulase enzyme production, predicting high plant tissue colonization abilities. Cellulase, crucial for breaking down cellulose, is essential for both industry and the environment. We sought to assess the potential of these bacteria as biocontrol agents against plant pathogenic fungi, with a focus on tomato plants. Bacterial isolates from insect bodies, including Lactococcus lactis, Pantoea ananatis, and Serratia liquefaciens, exhibited robust antifungal properties and cellulase activity. In vitro tests and glasshouse tests, confirmed their ability to inhibit the growth of plant-pathogenic fungi, indicating potential for biological control. Moreover, selected strains demonstrated high cellulase enzyme activity, vital for nutrient competition and rapid colonization of plant surfaces. The study introduces insect-gut-derived bacteria as promising biocontrol agents against plant pathogenic fungi. The identified strains, capable of inhibiting fungal growth and producing cellulase, offer sustainable alternatives to synthetic fungicides for protecting tomato plants. The findings advance agricultural practices by harnessing insect-associated bacteria, contributing to eco-friendly and efficient pest management strategies in modern agriculture.

Frailty and sarcopenia metabolomic signatures in kidney transplant candidates: the frailmar study

Abstract Sarcopenia and frailty are often overlooked in assessing kidney transplant (KT) candidates with chronic kidney disease (CKD), potentially leading to poor post-transplant outcomes. This study aimed to identify metabolites associated with frailty and sarcopenia in KT-candidates from the FRAILMAR study. Between June 2016 and June 2020, we evaluated frailty and sarcopenia in 173 KT-candidates using the Physical Frailty Phenotype and EGWSOP-2 criteria, respectively. Seventy-five metabolic markers from targeted pathways, previously linked to CKD, sarcopenia, or frailty, were measured in serum samples. These markers were analyzed using adjusted and weighted generalized linear models. Metabolomic data were integrated with multi-modal data, such as comorbidities, using a factor-based integration algorithm to identify metabolic phenotypes. Increased metabolites related to energy metabolism and essential amino acids were associated with frailty, mainly Krebs cycle intermediates. Sarcopenic KT-candidates showed lower levels of aromatic amino acids, lower protein/muscle metabolism, energy metabolism, and neurotransmission compared to non-sarcopenic patients. Unsupervised multi-modal integration revealed a high-risk metabolic phenotype characterized by the presence of sarcopenia, diabetes mellitus, and low body mass index, with alterations in branched-chain amino acids and high activity of lactate dehydrogenase enzyme. Frailty and sarcopenia are common among KT-candidates, and their metabolic status reveal notable disruptions in energy and amino acid metabolism. These findings highlight the value of a detailed metabolic assessment to more accurately evaluate patient health status prior to transplantation.

Effect of light intensity on nitrogen removal, enzymatic activity and metabolic pathway of algal-bacterial symbiosis in rotating biological contactor treating mariculture wastewater.

An algal-bacterial symbiosis (ABS) system was developed on a rotating biological contactor treating mariculture wastewater, and its nitrogen removal, enzymatic activity and metabolic pathways were investigated under different light intensities. The nitrogen removal efficiency increased when light intensities ranged from 20 to 80 μE/(m2·s) but declined under 100 μE/(m2·s). Higher enzymatic activities under 80 μE/(m2·s) facilitated nitrogen conversion, light utilization, ATP supply and photosynthesis. Reactive oxygen species accumulation activated antioxidant pathways under 20 and 100 μE/(m2·s). Functional bacteria including Sedimentitalea, Thauera and Dechloromonas as well as Chlorella sorokinian, Dunaliella, Pleurosira laevis and Microcystis were enriched under 80 μE/(m2·s). Abundant photosynthesis-related genes (petC, Lca3/4 and atpH/A) supported energy supply and electron transport. Conversely, lower proportions of IDH3, gltB, and acnA/B under 20 and 100 μE/(m2·s) hindered tricarboxylic acid cycle, reducing NADPH and energy production. These results enhance the understanding on the effect of light intensity on ABS system treating mariculture wastewater.

ZIF-8 nanocarriers synthesized by co-encapsulating resveratrol and cellulase for biomedical applications

Drug conjugation with enzymes is one of the innovative antibacterial nanocarriers used as a delivery system for cancer therapy. Zeolitic imidazolate framework-8 (ZIF-8) was synthesized, dual encapsulated with cellulase (CL) enzyme, and resveratrol (Resv) drug formed ZIF-8@CL&Resv. Cellulase and resveratrol hydrophobic nature have bound them together and imparted a negative charge on the ZIF-8, resulting in the decrease of zeta potential from 22.7 mV (ZIF-8) to 3.82 mV (ZIF-8@CL&Resv). The cellulase like a scaffold regulated the pH-responsive release of resveratrol enhancing its bioavailability. Molecular docking studies provided evidence of the major interaction between the biofilm-related proteins with cellulase and resveratrol. The encapsulated cellulase showed high enzymatic activity and possibly exhibited antibacterial effects by dissolving the biofilm and exposing bacteria to resveratrol action. Resveratrol released sustainably exhibited significant antioxidant and antibacterial activity against selected bacterial species. ZIF-8@CL&Resv exhibited high biocompatibility and had a potent cytotoxic effect against triple-negative breast cancer cells MDA MB 231 with an IC50-value of 17.18 μg/mL compared to ZIF-8 control with 90.47 μg/mL. ZIF-8@CL&Resv treatment led to 61.81 % cell death, apoptosis induction, increased ROS generation, and decreased mitochondrial membrane potential. Overall, data demonstrated that ZIF-8@CL&Resv is a novel drug release system and a potential catalytic nanoparticle for antimicrobial and anticancer applications.

Bacterial Degradation of Ibuprofen: Insights into Metabolites, Enzymes, and Environmental Fate Biodegradation of Ibuprofen by Achromobacter Species

In recent years, pharmaceuticals have emerged as pollutants due to their incomplete degradation in sewage treatment plants and their ability to cause physiological problems in humans even at low doses. Understanding the environmental fate of pharmaceutical pollutants and the mechanisms involved in their degradation is crucial for developing strategies to mitigate their impact on ecosystems and human health. In this study, the degradation of pharmaceutical compound ibuprofen was achieved by employing two bacterial strains, Achromobacter spanius strain S11 and Achromobacter piechaudii S18, previously isolated from contaminated water. These strains were capable of degrading ibuprofen as their sole carbon source. The study aimed to identify intermediate metabolites, determine the enzymes involved, and detect specific genes related to ibuprofen degradation. Different concentrations of ibuprofen, temperatures, and pH levels were tested. Both A. spanius S11 and A. piechaudii S18 successfully degraded ibuprofen. A. spanius S11 showed a degradation efficiency of 91.18% after only 72 h and reached 95.7% after 144 h, while A. piechaudii S18 exhibited degradation efficiencies of 72.39% and 73.01% after three and seven days, respectively. The LC-MS technique was used to identify biodegradation metabolites produced by A. spanius S11. The results indicated that the first step was hydroxylation followed by oxidation via the combination of monooxygenases that catalyze the C-H hydroxylation and dehydrogenases. Furthermore, the detection of intermediate metabolites of trihydroxyibuprofen suggested that the biodegradation of ibuprofen by A. spanius S11 can occur through multiple mechanisms. The highest enzyme activities were recorded for catechol 1,2-dioxygenase, 4.230 ± 0.026 U/mg, followed by laccase, 2.001 ± 0.215 U/mg. This study demonstrates the potential of Achromobacter strains, particularly A. spanius (S11), in degrading ibuprofen. These findings provide insights into the ibuprofen degradation process, intermediate metabolites, and relevant genes.

Potential of Indonesian cocoa pod husk for production of ligninolytic enzymes laccase and manganese peroxidase from Trametes hirsuta

Lignocellulosic biomass waste is a great potential source of cellulose and cellulose derivative bioproducts. Biomass pretreatment which involves ligninolytic enzymes is an environmentally friendly process that minimizes the use of harsh chemicals and energy input. This study evaluated white rot fungi and inducers (corncob, pineapple leaf, sorghum, and cocoa pod husk) to produce laccase (Lac) and manganese peroxidase (MnP). Enzyme production was optimized by using Plackett Burman and response surface methodology. Purification was undertaken by ammonium sulfate precipitation, dialysis, and anion exchange chromatography. Trametes hirsuta and cocoa pod husk as producer and inducer, respectively, resulted in the highest enzyme activities. The activities of Lac and MnP were optimum at 762.25 U/L and 596.92 U/L, respectively. The optimum pH and temperature for Lac and MnP were pH 3 (room temperature) and 60 °C (pH 5). The expected Lac and MnP were observed at ~65 kDa and 35–50 kDa protein band, respectively.

Screening and Identification of Industrial Enzyme Producing Bacteria from Sheep and Goat Milk

Industrial enzyme producing bacteria are essential for advancements in biotechnology. This study aims to identify bacterial strains from sheep and goat milk capable of producing industrially valuable enzymes. Basic Gram’s Staining and motility tests were performed from the bacterial colony of samples. Milk samples yielded a colony of bacteria which were screened for protease and amylase production. IMVIC tests demonstrated the feasibility of utilizing sheep and goat milk as a rich source of industrially valuable enzyme producing bacteria. By using SDS – PAGE the presence of the protein molecules is found. This study contributes to the discovery of novel enzyme-producing bacteria from unconventional sources, providing opportunities for the development of sustainable and efficient industrial processes. In this case, the results suggest that sheep and goat milk can serve as a rich source of industrially relevant enzymeproducing bacteria, offering potential applications in food processing, textile, and pharmaceutical industries. This demonstrated that certain bacterial strains exhibited high enzyme activity, highlighting their potential for industrial applications. Further studies on optimizing enzyme production and understanding the genetics of these isolates could lead to the development of new microbial strains for efficient enzyme production in various industries.

Protein Engineering of Nicotinamide Riboside Kinase Based on a Combinatorial Semirational Design Strategy for Efficient Biocatalytic Synthesis of Nicotinamide Mononucleotides.

Industrial biosynthesis of β-nicotinamide mononucleotide (β-NMN) lacks a highly active nicotinamide riboside kinase for the phosphorylation process. Cumbersome preprocessing steps and excessive ATP addition contribute to its increased cost. To tackle these challenges, a docking combination simulation (DCS) semirational mutagenesis strategy was designed in this study, combining various modification strategies to obtain a mutant NRK-TRA with 2.9-fold higher enzyme activity. Molecular dynamics simulations and structural analysis demonstrate the enhancement of its structural stability. High-density fermentation was achieved through a 5 L fermentation tank, with a titer reaching 208.3 U/mL, the highest in the current report. An ATP-cycling whole-cell catalytic system was employed and optimized by introducing a polyphosphate kinase 2 (PPK2) recombinant strain, and 15.16 g/L β-NMN was obtained through a series of batch transformation experiments. This study provides a new strategy for the efficient screening of highly active enzyme variants and offers a green and promising biotransformation system for NMN production.

Redox-mediated dsDNA-dye photooxidase mimic enable catalytic oxidation of 3,3′,5,5′-tetramethylbenzidine by dissolved O2 at neutral pH for improved biosensing

Catalytic oxidation of 3,3′,5,5′-Tetramethylbenzidine (TMB, an excellent chromogenic substrate) at neutral pH is critically important for amplified bioanalysis. Although some nanozymes exhibited the peroxidase activity at neutral pH, it is difficult to modulate their activity for homogeneous detection of biomolecules. In this work, we developed a redox-mediated dsDNA-dye photooxidase mimic that enables catalytic oxidation of TMB by dissolved O2 at neutral pH for improved biosensing. During illumination, the double-stranded DNA-SYBR Green I (dsDNA-SG) photogenerated singlet oxygen (1O2) can oxidized Mn2+ to Mn3+ that can efficiently oxidize TMB to produce a distinct blue within 4 min under neutral conditions. The catalytic oxidation of TMB can be readily modulated by the formation or dissociation of dsDNA during the sensing. After investigating a series of redox mediators, we found that only the Mn3+/Mn2+ redox mediator can lead to the oxidation of TMB at neutral pH. The maximum reaction rate of Mn2+-mediated dsDNA-SG photooxidase mimic under neutral conditions (pH 7.0) was 1.7 × 10−4 mM/s, even higher than that of horseradish peroxidase (HRP, 8.0 × 10−5 mM/s). The redox-mediated dsDNA-SG photooxidase mimic was used for detection of APE1 at pH 7.0 with over 130-fold higher sensitivity than that at 4.0, owing to the high enzymatic activity of APE1 at neutral pH. Meanwhile, we further extended this photooxidase mimic for the sensitive detection of DNA (LOD, 8 pM) and heavy metal ions at neutral pH. The redox-mediated dsDNA-dye photooxidase mimic with the ease of modulating its enzymatic activity and working at neutral pH is quite appealing for biosensing.

Direct Preparation of Alginate Oligosaccharides from Brown Algae by an Algae-Decomposing Alginate Lyase AlyP18 from the Marine Bacterium Pseudoalteromonas agarivorans A3.

Alginate oligosaccharides (AOs), derived from alginate degradation, exhibit diverse biological activities and hold significant promise in various fields. The enzymatic preparation of AOs relies on alginate lyases, which offers distinct advantages. In contrast to the conventional use of sodium alginate derived from brown algae as the substrate for the enzymatic preparation of AOs, AO preparation directly from brown algae is more appealing due to its time and energy efficiency. Thus, the identification of potent alginate lyases and cost-effective brown algae substrates is crucial for optimizing AO production. Herein, we identified and characterized an alginate lyase, AlyP18, capable of efficiently decomposing algae, from a marine bacterium Pseudoalteromonas agarivorans A3 based on secretome analysis. AlyP18 is a mesothermal, endo-type and bifunctional alginate lyase with high enzymatic activity. Two brown algae substrates, Laminaria japonica roots and Macrocystis pyrifera, were used for the AO preparation by AlyP18. Upon optimization of AlyP18 hydrolysis parameters, the substrate degradation efficiency and AO production reached 53% and ~32% for L. japonica roots, respectively, and 77% and ~46.5% for M. pyrifera. The generated AOs primarily consisted of dimers to pentamers, with trimers and tetramers being dominant. This study provides an efficient alginate lyase and alternative brown algal feedstock for the bioconversion of high-value AOs from brown algae.

Reductive dehalogenase of Dehalococcoides mccartyi strain CBDB1 reduces cobalt- containing metal complexes enabling anodic respiration.

Microorganisms capable of direct or mediated extracellular electron transfer (EET) have garnered significant attention for their various biotechnological applications, such as bioremediation, metal recovery, wastewater treatment, energy generation in microbial fuel cells, and microbial or enzymatic electrosynthesis. One microorganism of particular interest is the organohalide-respiring bacterium Dehalococcoides mccartyi strain CBDB1, known for its ability to reductively dehalogenate toxic and persistent halogenated organic compounds through organohalide respiration (OHR), using halogenated organics as terminal electron acceptors. A membrane-bound OHR protein complex couples electron transfer to proton translocation across the membrane, generating a proton motive force, which enables metabolism and proliferation. In this study we show that the halogenated compounds can be replaced with redox mediators that can putatively shuttle electrons between the OHR complex and the anode, coupling D. mccartyi cells to an electrode via mediated EET. We identified cobalt-containing metal complexes, referred to as cobalt chelates, as promising mediators using a photometric high throughput methyl viologen-based enzyme activity assay. Through various biochemical approaches, we show that cobalt chelates are specifically reduced by CBDB1 cells, putatively by the reductive dehalogenase subunit (RdhA) of the OHR complex. Using cyclic voltammetry, we also demonstrate that cobalt chelates exchange electrons with a gold electrode, making them promising candidates for bioelectrochemical cultivation. Furthermore, using the AlphaFold 2-calculated RdhA structure and molecular docking, we found that one of the identified cobalt chelates exhibits favorable binding to RdhA, with a binding energy of approximately -28 kJ mol-1. Taken together, our results indicate that bioelectrochemical cultivation of D. mccartyi with cobalt chelates as anode mediators, instead of toxic halogenated compounds, is feasible, which opens new perspectives for bioremediation and other biotechnological applications of strain CBDB1.

Degradation of High Concentrations of Anthracene Using White-Rot Wood-Inhabiting Fungi and Investigation of Enzyme Activities

Owing to the production of lignin-modifying enzymes (LMEs), white-rot fungi (WRF) such as polypores are potent organisms in the biodegradation of xenobiotic pollutants. The nonspecific function of LMEs including laccase and manganese peroxidase (MnP), has enabled the use of WRF in biotechnological applications, particularly in bioremediation. In this study, 12 strains from nine white-rot basidiomycete genera viz., Ganoderma, Inocutis, Irpex, Lentinus, Lenzites, Oxyporus, Peniophora, Sanghuangporus, and Trametes were isolated from Iran and identified using morphological and molecular tools. The enzyme activity of laccase and manganese peroxidase that directly correlated with the biodegradation were determined, and the strains with the highest enzyme activities were evaluated for their ability to degrade 400 mg/L of anthracene over 28 days. Gas chromatography with flame ionization detector (GC-FID) revealed that four polypores viz., Trametes versicolor v21te, T. versicolor v22da, T. hirsuta, and Oxyporus sp. degraded 64%, 52%, 34%, and 20% of the anthracene, respectively. According to our analysis, the examined indigenous WRF are potentially useful candidates for the development of new mycoremediation techniques to degrade polycyclic aromatic hydrocarbons (PAHs).

Exploring the potential of triticale lines for bioethanol production

Aim: Triticale is a well adaptable crop, tolerant of disease and abiotic stresses, and able to grow with good yields even in poor soil, thus representing a good choice to develop a new industrial agri-chain in Italy in a sustainability contest, to cope with its soil problems due to incoming desertification. Methods: Two triticale elite lines were grown in marginal lands in controlled field experiments. The lines were harvested at two different development stages, namely green mass and seeds, and suitable standard protocols were applied to test their potential to produce bioethanol in line with the emerging bioenergy processes. Results: The protocols applied were able to obtain bioethanol with a good yield from both feedstocks. In particular, very efficient fermentation kinetics was observed using seed feedstock, with a sharp curve between 15 h and 24 h, reaching 84% of the total alcohol obtained (final time 72 h). Conclusions: Therefore, the results of this research point to new sustainable potential for industrial applications of triticale crops in Italy. Furthermore, the high activity of the endogenous amylolytic enzymes, mainly α-amylase, and the high starch content suggest the potential use of triticale in other industrial applications, like the brewing industry.

Long term impact of manuring, fertilization and liming on soil resilience, enzyme activities and productivity under maize-wheat system in an acidic Alfisol

Aim: This study was carried out to evaluate the long-term impact of manuring, fertilization and liming on soil resilience, enzyme activities and system productivity under changing climate scenarios. Methodology: The soil samples were collected from 67 year old long-term experiment going on, at Birsa Agricultural University, Jharkhand during 2022-23 under maize-wheat cropping system from seven treatments, viz. T1: control, T2: 100 % N, T3: 100 % FYM, T4: ½ (N+FYM) + PX/2 + KY/2, T5: 100 % NPK, T6: Lime + NPK, T7: Lime + FYM +P(A-X) + K(B-Y). To assess soil resistance and resilience, surface samples were subjected to with and without heat stress (48±2°C for 80 hrs) and substrate addition (0.02 g glucose g-1 soil). Results: Application of FYM/lime with NPK significantly improved bulk density, pH, SOC and system productivity. The highest enzyme activities and cumulative respiration were observed in lime + FYM + P(A-X) + K(B-Y) treatment. It exhibited the highest resistance index (0.92) and resilience index (0.34). Interpretation: The integrated use of inorganic, organic along with liming proved to be a useful management strategy for maintaining soil quality, resistance and resilience, and crop productivity in acid soil of Jharkhand. Key words: Enzyme, Heat stress, Maize-wheat system, Soil acidification, Substrate induced respiration, Wheat-maize

Isolation and identification of cellulose decomposition bacteria from the soil of Yunnan province, China

In this experiment sodium carboxymethyl cellulose (CMC-Na) was used as the only carbon source to isolate cellulose decomposing bacteria from the soil of Qiongzhusi Forest Park in Kunming, Yunnan Province. The ratio of hydrolytic circle diameter and colony diameter formed by the strain on cellulose Congo red medium was determined for primary screening, and then the filter paper disintegration test and CMC enzyme activity assay were used for rescreen. A cellulose decomposing bacterium (HS-2) with high enzyme activity was obtained, and its enzyme activity was up to 16.42 U/ml. (U means the unit of activity of enzymes, activity, unit) Gram stain and Spore stain reaction showed that HS-2 strain was Gram-positive Bacillus. The Bacillus strain was identified as Bacillus pumilus on the basis of physiological and biochemical tests, and 16S rDNA sequence analysis. Bangladesh J. Bot. 52(3): 635-641, 2024 (September) Special