Optimization of cold-active CMCase production by psychrotrophic Sphingomonas sp. FLX-7 from the cold region of China

Li, Dapeng; Feng, Lei; Liu, Keran; Cheng, Yi; Hou, Ning; Li, Chunyan

doi:10.1007/s10570-016-0859-4

Cited by 26 publications

(14 citation statements)

References 34 publications

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“…CMCase (endoglucanase) activity was determined by the 3,5-dinitrosalicyclic acid (DNS) colorimetric method as described by Ariffin et al (2006) and Li et al (2016). The crude enzyme (0.5 ml) was mixed with 2 ml of substrate solution (1% w/v CMC dissolved in 50 mM sodium citrate buffer (pH 4.8)) and incubated for 30 min at 30°C in a water bath.…”

Section: Enzyme Assaysmentioning

confidence: 99%

“…Although it is typically assumed that fungi are the major decomposers of complex plant biopolymers (Boer et al 2005), numerous bacteria have been described in recent years as efficient cellulose degraders (de Cassia Pereira et al 2017;Banerjee et al 2020), often more effective than fungi (Ariffin et al 2006). Bacteria have a number of advantages compared to fungi: higher growth rates allow for the higher recombinant production of enzymes, they express multienzyme complexes with increased function and synergy, and they inhabit a wide variety of environments and are often resistant to extreme conditions (Sadhu and Maiti 2013;Li et al 2016).…”

Section: Introductionmentioning

confidence: 99%

“…Most studies on cellulolytic bacteria have been focused on mesophilic or thermophilic representatives. The number of studies on cellulose degraders in cold environments is low (Li et al 2016;Lin et al 2019;Sun et al 2020). It has been recognized that cold-active enzymes provide a wide biotechnological potential.…”

Section: Introductionmentioning

confidence: 99%

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Production and partial characterization of a crude cold-active cellulase (CMCase) from Bacillus mycoides AR20-61 isolated from an Alpine forest site

Steiner

Margesin

2020

Ann Microbiol

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Purpose To evaluate the production of a cold-active CMCase (endoglucanase) by Bacillus mycoides AR20-61 isolated from Alpine forest soil and to characterize the crude enzyme. Methods After studying the effect of cultivation parameters (medium composition, temperature, NaCl concentration, pH) on bacterial growth and enzyme production, the crude enzyme was characterized with regard to the effect of pH, temperature, and inhibitors on enzyme activity and stability. Result Optimum growth and enzyme production occurred at 20–25 °C, pH 7, and 1–1.5% (w/v) CMC. Despite high biomass production over the whole growth temperature range (10–35 °C), enzyme production was low at 10 and 35 °C. CMC concentration had a minor effect on growth, independent of the growth temperature, but a significant effect on CMCase production at temperatures ≥ 20 °C. The crude enzyme was active over a broad temperature range (0–60 °C); the apparent optimum temperature for activity was at 40–50 °C. The cultivation temperature influenced the effect of temperature on enzyme activity and stability. A significantly higher thermosensitivity of the enzyme produced at a cultivation temperature of 10 °C compared to that produced at 25 °C was noted at 50 and 65 °C. The enzyme was highly active over a pH range of 4–6 and showed optimum activity at pH 5. No activity was lost after 60 min of incubation at 30 °C and pH 4–9. The CMCase was resistant against a number of monovalent and divalent metal ions, metal-chelating agents, and phenol. Conclusion The CMCase produced by the studied strain is characterized by high activities in the low temperature range (down to 0 °C) and acidic pH range, high stability over a broad pH range, and high resistance against a number of effectors. Our results also demonstrate the different, independent roles of temperature in bacterial growth, enzyme production, nutrient requirements during enzyme production, and enzyme characteristics regarding thermosensitivity, which has not yet been described for cellulases.

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Section: Enzyme Assaysmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Production and partial characterization of a crude cold-active cellulase (CMCase) from Bacillus mycoides AR20-61 isolated from an Alpine forest site

Steiner

Margesin

2020

Ann Microbiol

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“…These data suggest that the strongly labeled cellulolytic strain is degrading 13 C-cellulose extracellularly and the weakly 13 C-enriched strain can access degradation products as well as other sources of unlabeled carbon present in soil. The capacity of Sphingomonas species to degrade cellulose through the activity of extracellular enzymes is known [106,107].…”

Section: Mutualists Opportunists and Parasitesmentioning

confidence: 99%

Competitive exclusion and metabolic dependency among microorganisms structure the cellulose economy of agricultural soil

Wilhelm¹,

Pepe-Ranney

Weisenhorn

et al. 2020

Preprint

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Many cellulolytic microorganisms degrade cellulose through extracellular processes that yield free intermediates which promote interactions with non-cellulolytic organisms. We hypothesize that these interactions determine the ecological and physiological traits that govern the fate of cellulosic carbon (C) in soil. We evaluated the genomic potential of soil microorganisms that access C from 13 C-labeled cellulose. We used metagenomic-SIP and metaproteomics to evaluate whether cellulolytic and non-cellulolytic microbes that access 13 C from cellulose encode traits indicative of metabolic dependency or competitive exclusion. The most highly 13 C-enriched taxa were cellulolytic Cellvibrio ( Gammaproteobacteria ) and Chaetomium ( Ascomycota ), which exhibited a strategy of self-sufficiency (prototrophy), rapid growth, and competitive exclusion via antibiotic production. These ruderal taxa were common indicators of soil disturbance in agroecosystems, such as tillage and fertilization. Auxotrophy was more prevalent in cellulolytic Actinobacteria than in cellulolytic Proteobacteria , demonstrating differences in dependency among cellulose degraders. Non-cellulolytic taxa that accessed 13 C from cellulose ( Planctomycetales , Verrucomicrobia and Vampirovibrionales ) were highly dependent, as indicated by patterns of auxotrophy and 13 C-labeling (i.e. partial labelling or labeling at later-stages). Major 13 C-labeled cellulolytic microbes ( e.g. Sorangium, Actinomycetales, Rhizobiales and Caulobacteraceae ) possessed adaptations for surface colonization ( e.g. gliding motility, hyphae, attachment structures) signifying the importance of surface ecology in decomposition. These results suggest that access to cellulose was accompanied by ecological trade-offs characterized by differing degrees of metabolic dependency and competitive exclusion. These trade-offs likely influence microbial growth dynamics on particulate organic carbon and reveal that the fate of carbon is governed by a complex economy within the microbial community.

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Section: Mutualists Opportunists and Parasitesmentioning

confidence: 99%

Competitive exclusion and metabolic dependency among microorganisms structure the cellulose economy of agricultural soil

Wilhelm

Pepe-Ranney

Weisenhorn

et al. 2020

Preprint

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Background Microorganisms that degrade cellulose utilize extracellular processes that yield free intermediates which promote interactions with non-cellulolytic organisms. We hypothesized that these interactions determine the ecological and physiological traits governing the fate of cellulosic carbon (C) in soil. We used data from a metagenomic-SIP experiment to perform comparative genomics and characterize the attributes of cellulolytic and non-cellulolytic taxa accessing 13C from cellulose. We hypothesized that cellulolytic taxa would exhibit competitive traits to limit access, while non-cellulolytic taxa would display metabolic dependency, such as signatures of adaptive gene loss. We tested our hypotheses by evaluating genomic traits indicative of competitive exclusion or metabolic dependency, such as antibiotic production, growth rate, surface attachment, biomass degrading potential and auxotrophy.Results The most 13C-enriched taxa were cellulolytic Cellvibrio (Gammaproteobacteria) and Chaetomium (Ascomycota), which exhibited a strategy of self-sufficiency (prototrophy), rapid growth, and competitive exclusion via antibiotic production. Auxotrophy was more prevalent in cellulolytic Actinobacteria than in cellulolytic Proteobacteria, demonstrating differences in dependency among cellulose degraders. Non-cellulolytic taxa that accessed 13C from cellulose (Planctomycetales, Verrucomicrobia and Vampirovibrionales) were highly dependent, as indicated by patterns of auxotrophy and 13C-labeling (i.e. partial labelling or labeling at later-stages). Major 13C-labeled cellulolytic microbes (e.g. Sorangium, Actinomycetales, Rhizobiales and Caulobacteraceae) possessed adaptations for surface colonization (e.g. gliding motility, hyphae, attachment structures) signifying the importance of surface ecology in decomposition. Conclusions Our results demonstrate that access to cellulose was accompanied by ecological trade-offs characterized by differing degrees of metabolic dependency and competitive exclusion. These trade-offs influence microbial growth dynamics on particulate organic carbon and reveal that the fate of carbon is governed by a complex economy within the microbial community. We propose three ecological groups to describe participants in this economy: (i) independent primary degraders, (ii) integrated primary degraders and (iii) mutualists, opportunists and parasites. The relative importance and taxonomic composition of these groups reported here should be considered context dependent, likely reflecting disturbance and management practices common to agricultural soils.

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Optimization of cold-active CMCase production by psychrotrophic Sphingomonas sp. FLX-7 from the cold region of China

Cited by 26 publications

References 34 publications

Production and partial characterization of a crude cold-active cellulase (CMCase) from Bacillus mycoides AR20-61 isolated from an Alpine forest site

Production and partial characterization of a crude cold-active cellulase (CMCase) from Bacillus mycoides AR20-61 isolated from an Alpine forest site

Competitive exclusion and metabolic dependency among microorganisms structure the cellulose economy of agricultural soil

Competitive exclusion and metabolic dependency among microorganisms structure the cellulose economy of agricultural soil

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