API ZYM assay to evaluate enzyme fingerprinting and microbial functional diversity in relation to soil processes

Martinez, Diego Stéfani T.; Molina, Marirosa; Sánchez, Joaquín Fernández; Moscatelli, M.C.; Marinari, Sara

doi:10.1007/s00374-015-1055-7

Cited by 22 publications

(27 citation statements)

References 31 publications

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“…Enzyme fingerprinting using APIZYM assay. The relative activities for soil enzyme were measured by a semi-quantitative APIZYM system (Biomerieus, USA) according to the manufacturer's protocol 16,46,47 . The APIZYM kit supplied with 20 microcupules, contains dehydrated chromogenic substrates for different hydrolytic enzymes and control ( Supplementary Table S5).…”

Section: Methodsmentioning

confidence: 99%

Steel slag amendment impacts on soil microbial communities and activities of rice (Oryza sativa L.)

Das

Gwon

Khan

et al. 2020

Sci Rep

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With the increase in iron/steel production, the higher volume of by-products (slag) generated necessitates its efficient recycling. Because the Linz-Donawitz (LD) slag is rich in silicon (Si) and other fertilizer components, we aim to evaluate the impact of the LD slag amendment on soil quality (by measuring soil physicochemical and biological properties), plant nutrient uptake, and strengthens correlations between nutrient uptake and soil bacterial communities. We used 16 S rRNA illumine sequencing to study soil bacterial community and APIZYM assay to study soil enzymes involved in C, N, and P cycling. The LD slag was applied at 2 Mg ha −1 to Japonica and indica rice cultivated under flooded conditions. The LD slag amendment significantly improved soil pH, plant photosynthesis, soil nutrient availability, and the crop yield, irrespective of cultivars. It significantly increased N, P, and Si uptake of rice straw. The slag amendment enhanced soil microbial biomass, soil enzyme activities and enriched certain bacterial taxa featuring copiotrophic lifestyles and having the potential role for ecosystem services provided to the benefit of the plant. The study evidenced that the short-term LD slag amendment in rice cropping systems is useful to improve soil physicochemical and biological status, and the crop yield. Soil enzyme activities.Labile carbon degrading enzymes such as α-glucosidase, β-glucosidase, α-galactosidase, β-galactosidase, α-mannosidase, and α-fucosidase which can degrade maltose, cellobiose, melibiose, lactose, mannose, and fructose, respectively, markedly increased in the slag amendment treatments in comparison to the unamended treatments, irrespective of rice cultivars (Fig. 1). Likewise, recalcitrant carbon degrading enzymes such as esterase, lipase, and N-acetyl-β-glucosaminidase which can degrade hemicelluloses and polysaccharide respectively, increased by the slag amendment. Notably, the increase was more in labile carbon degrading enzymes (mostly β-glucosidase, and β-galactosidase) compared to the recalcitrant carbon degrading enzymes. The LD slag amendment also noticeably increased soil enzyme activities involved in nitrogen cycling (i.e., leucine-aminopeptidase and trypsin) and phosphorus cycling (i.e., alkaline phosphomonoesterase and phosphohydrolase). Notably, the increase in soil enzyme activities was more prominent in Indica in comparison to Japonica rice variety. Among the C cycling enzyme, β-glucosidase, and β-galactosidase were dominant, whereas among the N cycling enzyme, aminopeptidase and among the P cycling enzyme, alkaline phosphomonoesterase were dominant in the paddy soil (Fig. 1).Relative abundance and diversity of rhizosphere bacterial community. The most abundant (≥ 1.0%) bacterial phyla across treatments were Proteobacteria (34-48%), Fermicutes (15-17%), Actinobacteria (7-18%), Acidobacteria (3-7%), Bacteroidetes (2-6%), Nitrospirae (2-3%), and Chloroflexi (1-4%). The LD slag amendment significantly (P < 0.05) increased the relative abundance of Proteobacteria by 28.9 and 2...

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Section: Methodsmentioning

confidence: 99%

Steel slag amendment impacts on soil microbial communities and activities of rice (Oryza sativa L.)

Das

Gwon

Khan

et al. 2020

Sci Rep

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“…After this time, the enzyme activity intensity reading was taken by assigning numerical values from 0 to 5 to each reaction according to the manufacturer's reading table and reported in the literature [34,36,37]. Colour intensity readings for enzymatic reactions were performed by 4 independent observers [38]. With the results obtained, an extracellular enzyme profile diagram was designed as established by Patel et al [39].…”

Section: Biological Characterisation and Enzymatic Profile Of Biosolidsmentioning

confidence: 99%

“…However, little information is available on the enzymatic activity of biosolids due to the lack of methodologies or analytical methods that provide information quickly and inexpensively. In recent decades, the API ZYM ® system has been used and validated to evaluate enzymatic profiles of soils, compost, activated sludge and sediments [34,[36][37][38][39]44,134,137,138]. However, the API ZYM ® system has not been used directly and over time in biosolids samples as a system for the rapid determination of 19 extracellular hydrolytic enzymatic reactions related to the C, N and P cycles.…”

Section: Extracellular Enzyme Profiles Per Api Zym ® Kit For Biosolidsmentioning

confidence: 99%

“…Also included are enzyme activity monitoring systems that allow the establishment of "fingerprints" or activity enzyme "profiles" such as the API ZYM ® system. As well, the establishment of indices of functional diversity and microbial activity, such as the Shannon index (H') and synthetic enzymatic index (SEI) is also important for their potential contribution as indicators of quality and agronomic functionality of biosolids before and after being applied as soil improvers [24][25][26][27][28][29][30][31][32][33][34][35][36][37][38][39][40]. Only U.S.A. regulation stipulates that the agronomic speed of application cannot be exceeded [41]; to protect the quality of groundwater or surface water, nitrogen must be regulated through an agronomic rate approach, which requires an estimate of the need for N from the crop and the availability of N in biosolids [41,42].…”

Section: Introductionmentioning

confidence: 99%

“…Several enzyme assays are available to determine the hydrolytic activity in different environmental matrices. One of these enzymatic assays is the API ZYM ® system which has been used in environmental matrices with complex biochemical processes e.g., soils, compost, activated sludge and sediments [34][35][36][37][38][39]44]. This system provides rapid and semi-quantitative information on the hydrolytic activity of 19 enzymes associated with the biodegradation of lipids, proteins, carbohydrates, and nucleic acids [34,36,39].…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of Non-Conventional Biological and Molecular Parameters as Potential Indicators of Quality and Functionality of Urban Biosolids Used as Organic Amendments of Agricultural Soils

et al. 2020

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Biosolids are waste from wastewater treatment and have a high content of organic matter and nutrients. In this study, not conventional physicochemical and biological properties of biosolids produced during different seasons of the year were evaluated. These properties are not considered in environmental regulations; however, they are of agronomic interest as indicators of quality and functionality in soils. Also, molecular analysis by Fourier-transform infrared (FTIR) was conducted, enzymatic analysis using the APIZYM® system was performed and two indices of functional and microbial diversity were established. The results showed that the biosolids had a high content of total organic carbon, total nitrogen, P, and K. FTIR analyses showed that chemical composition of biosolids was similar during all year. The C and N of microbial biomass demonstrated presence of active microorganisms, as well as a uniformity in its richness and abundance of species that could present a positive synergy with soil microorganisms. The enzymatic activities showed that the biosolids contained an enzymatic machinery available to promote the mineralisation of the organic matter of biosolids and could even transcend into the soil. Finally, biological properties can be used as indicators of quality and functionality of biosolids before being used as an organic amendment, especially in agricultural soils.

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Characterisation of recombinant GH 3 β-glucosidase from β-glucan producing Levilactobacillus brevis TMW 1.2112

Bockwoldt

Ehrmann

2022

Antonie van Leeuwenhoek

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Levilactobacillus (L.) brevis TMW 1.2112 is an isolate from wheat beer that produces O2-substituted (1,3)-β-D-glucan, a capsular exopolysaccharide (EPS) from activated sugar nucleotide precursors by use of a glycosyltransferase. Within the genome sequence of L. brevis TMW 1.2112 enzymes of the glycoside hydrolases families were identified. Glycoside hydrolases (GH) are carbohydrate-active enzymes, able to hydrolyse glycosidic bonds. The enzyme β-glucosidase BglB (AZI09_02170) was heterologous expressed in Escherichia coli BL21. BglB has a monomeric structure of 83.5 kDa and is a member of the glycoside hydrolase family 3 (GH 3) which strongly favoured substrates with β-glycosidic bonds. Km was 0.22 mM for pNP β-D-glucopyranoside demonstrating a high affinity of the recombinant enzyme for the substrate. Enzymes able to degrade the (1,3)-β-D-glucan of L. brevis TMW 1.2112 have not yet been described. However, BglB showed only a low hydrolytic activity towards the EPS, which was measured by means of the D-glucose releases. Besides, characterised GH 3 β-glucosidases from various lactic acid bacteria (LAB) were phylogenetically analysed to identify connections in terms of enzymatic activity and β-glucan formation. This revealed that the family of GH 3 β-glucosidases of LABs comprises most likely exo-active enzymes which are not directly associated with the ability of these LAB to produce EPS.

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API ZYM assay to evaluate enzyme fingerprinting and microbial functional diversity in relation to soil processes

Cited by 22 publications

References 31 publications

Steel slag amendment impacts on soil microbial communities and activities of rice (Oryza sativa L.)

Steel slag amendment impacts on soil microbial communities and activities of rice (Oryza sativa L.)

Evaluation of Non-Conventional Biological and Molecular Parameters as Potential Indicators of Quality and Functionality of Urban Biosolids Used as Organic Amendments of Agricultural Soils

Characterisation of recombinant GH 3 β-glucosidase from β-glucan producing Levilactobacillus brevis TMW 1.2112

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