Vector-initiated transitive RNA interference in the filamentous fungus Aspergillus oryzae

Fernandez, Evee Q.; Moyer, Donna L.; Maiyuran, Suchindra; Labaro, Anna; Brody, Howard

doi:10.1016/j.fgb.2012.01.011

Cited by 16 publications

(14 citation statements)

References 43 publications

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“…The purified PCR products were digested, reclaimed and cloned into plasmid pSKNHG, generating recombinant expression vector pSKNHG-AmyA1. Afterwards, PEG-CaCl 2 mediated transformation was completed as previously described [ 16 ]. Transformants were selected according to their growth capability on MM plates, and then confirmed by PCR.…”

Section: Methodsmentioning

confidence: 99%

Functional expression of a novel α-amylase from Antarctic psychrotolerant fungus for baking industry and its magnetic immobilization

Mao

Zhang

et al. 2017

BMC Biotechnol

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Backgroundα-Amylase plays a pivotal role in a broad range of industrial processes. To meet increasing demands of biocatalytic tasks, considerable efforts have been made to isolate enzymes produced by extremophiles. However, the relevant data of α-amylases from cold-adapted fungi are still insufficient. In addition, bread quality presents a particular interest due to its high consummation. Thus developing amylases to improve textural properties could combine health benefits with good sensory properties. Furthermore, iron oxide nanoparticles provide an economical and convenient method for separation of biomacromolecules. In order to maximize the catalytic efficiency of α-amylase and support further applications, a comprehensive characterization of magnetic immobilization of α-amylase is crucial and needed.ResultsA novel α-amylase (AmyA1) containing an open reading frame of 1482 bp was cloned from Antarctic psychrotolerant fungus G. pannorum and then expressed in the newly constructed Aspergillus oryzae system. The purified recombinant AmyA1 was approximate 52 kDa. AmyA1 was optimally active at pH 5.0 and 40 °C, and retained over 20% of maximal activity at 0–20 °C. The K m and V max values toward soluble starch were 2.51 mg/mL and 8.24 × 10−2 mg/(mL min) respectively, with specific activity of 12.8 × 103 U/mg. AmyA1 presented broad substrate specificity, and the main hydrolysis products were glucose, maltose, and maltotetraose. The influence of AmyA1 on the quality of bread was further investigated. The application study shows a 26% increase in specific volume, 14.5% increase in cohesiveness and 14.1% decrease in gumminess in comparison with the control. AmyA1 was immobilized on magnetic nanoparticles and characterized. The immobilized enzyme showed improved thermostability and enhanced pH tolerance under neutral conditions. Also, magnetically immobilized AmyA1 can be easily recovered and reused for maximum utilization.ConclusionsA novel α-amylase (AmyA1) from Antarctic psychrotolerant fungus was cloned, heterologous expression in Aspergillus oryzae, and characterized. The detailed report of the enzymatic properties of AmyA1 gives new insights into fungal cold-adapted amylase. Application study showed potential value of AmyA1 in the food and starch fields. In addition, AmyA1 was immobilized on magnetic nanoparticles and characterized. The improved stability and longer service life of AmyA1 could potentially benefit industrial applications.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-017-0343-8) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Functional expression of a novel α-amylase from Antarctic psychrotolerant fungus for baking industry and its magnetic immobilization

Mao

Zhang

et al. 2017

BMC Biotechnol

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“…In Caenorhabditis elegans , transitivity is only detected in the 5′ region of the original targeted site ( Sijen et al, 2001 ; Alder et al, 2003 ). In fungi, secondary siRNA has been reported to originate from either region, with transitivity occurring upstream for Schizosaccharomyces pombe and Aspergillus oryzae ( Simmer et al, 2010 ; Fernandez et al, 2012 ), or downstream of the targeted area for Mucor circinelloides ( Nicolás et al, 2003 ). In plants, transitivity is bi-directional, with the silencing signal spreading to both directions ( Vaistij et al, 2002 ).…”

Section: Transitivity Spreading Pattern In Plantsmentioning

confidence: 99%

The Whys and Wherefores of Transitivity in Plants

Felippes

Waterhouse

2020

Front. Plant Sci.

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Transitivity in plants is a mechanism that produces secondary small interfering RNAs (siRNAs) from a transcript targeted by primary small RNAs (sRNAs). It expands the silencing signal to additional sequences of the transcript. The process requires RNA-dependent RNA polymerases (RDRs), which convert single-stranded RNA targets into a double-stranded (ds) RNA, the precursor of siRNAs and is critical for effective and amplified responses to virus infection. It is also important for the production of endogenous secondary siRNAs, such as phased siRNAs (phasiRNAs), which regulate several genes involved in development and adaptation. Transitivity on endogenous transcripts is very specific, utilizing special primary sRNAs, such as miRNAs with unique features, and particular ARGONAUTEs. In contrast, transitivity on transgene and virus (exogenous) transcripts is more generic. This dichotomy of responses implies the existence of a mechanism that differentiates self from non-self targets. In this work, we examine the possible mechanistic process behind the dichotomy and the intriguing counter-intuitive directionality of transitive sequence-spread in plants.

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“…The rdrp-2 gene is required for accumulating secondary siRNAs and abundantly expressed in proportion to dsRNA production (Calo et al, 2012). In Aspergillus oryzae, an RdRP is necessary for transitive RNAi and expression correlates with transitive RNAi efficiency (Fernandez et al, 2012). On the other hand, RdRPs are not necessary for RNAi and 3 0 transitive RNAi was not observed in Aspergillus nidulans (Hammond and Keller, 2005).…”

Section: Introductionmentioning

confidence: 99%

Systemic RNA interference is not triggered by locally-induced RNA interference in a plant pathogenic fungus, Rosellinia necatrix

Shimizu

Yaegashi

Ito

et al. 2015

Fungal Genetics and Biology

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Vector-initiated transitive RNA interference in the filamentous fungus Aspergillus oryzae

Cited by 16 publications

References 43 publications

Functional expression of a novel α-amylase from Antarctic psychrotolerant fungus for baking industry and its magnetic immobilization

Functional expression of a novel α-amylase from Antarctic psychrotolerant fungus for baking industry and its magnetic immobilization

The Whys and Wherefores of Transitivity in Plants

Systemic RNA interference is not triggered by locally-induced RNA interference in a plant pathogenic fungus, Rosellinia necatrix

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