Karim Imangholiloo; Nasrin Moshtaghi; Seyed Hasan Marashi; Abdolreza Bagheri; Ahmad Sharifi
Abstract
Cellulose which is extremely produced by plants, can be used for biofuel production but this function needs chemical or enzymatic digestion. Cellulose hydrolysis of plant wastes for ethanol production requires a mixture of three enzyme groups, including endoglucanases, exoglucanases, and beta-glucosidases. ...
Read More
Cellulose which is extremely produced by plants, can be used for biofuel production but this function needs chemical or enzymatic digestion. Cellulose hydrolysis of plant wastes for ethanol production requires a mixture of three enzyme groups, including endoglucanases, exoglucanases, and beta-glucosidases. The cellobiohydrolase enzyme (Cel6B) from Thermobifidia fusca has been used for cellulase activity extensively. This research aimed to express recombinant Cel6B enzyme in Pichia pastoris. For this purpose, cel6B gene in control of AOX1 promoter (methanol inducible) was introduced into Pichia pastoris. Amplification of cel6B gene was performed by PCR technique and then introduced into the Phil-S1 yeast vector. The recombinant construct contained the cel6B gene sequence and PHO1 signal sequence as secretion signal was transferred into Pichia pastoris GS115 strain. The transformed yeast cells expressed the recombinant Cel6B to yield 2.104 U (µmol/min)/ml of culture medium. Purified recombinant enzyme showed the best activity at 60 °C and pH 4.5 and this was agreed with optimum conditions for recombinant Cel6B enzymes which were produced in other systems. The purity of the enzyme was examined by SDS–PAGE technique, and a single band with a molecular weight about 59.6 kDa was observed. As cel6B gene sequence was not optimized for expression in the Pichia pastoris yeast, this could be one of the reasons for low level activity of recombinant Cel6B enzyme. This thermostable enzyme can be used for cellulolytic digestion of biomaterials in biofuel production research and other uses.
Atieh Teymoori; Mojtaba Teimoori; Madjid Momeni-Moghaddam
Abstract
For 50 years, the term gene is synonymous with regions of the genome gene that coding by mRNAs and translate to protein. nonetheless, Genome wide Recent studies have revealed that regulating gene expression through degradation or translational inhibition of their point mRNAs and thus attend in a wide ...
Read More
For 50 years, the term gene is synonymous with regions of the genome gene that coding by mRNAs and translate to protein. nonetheless, Genome wide Recent studies have revealed that regulating gene expression through degradation or translational inhibition of their point mRNAs and thus attend in a wide variety of physiological and pathological cellular processes including: development, cell proliferation, differentiation, and apoptosis pathways by thousands of regulatory non coding RNA such as lncRNAs and microRNAs. According to a recent survey, it is known this RNAs have vital role in regulation cellular pathways at transcriptional, posttranscriptional and epigenetic levels. These noncoding genes are often aberrantly expressed in a variety of human cancers. However, the biological functions of most ncRNAs remain largely in doubt. In this review, we proved that a remarkable part of the genetic etiology of cancer is imposed by noncoding regulatory sequences. The purpose of this review is aimed to give an outlook of using of noncoding RNA as diagnostic markers and therapeutic targets. These observations emphasized that the recognition of coding genes and Research continued evolution and function of non-coding RNAs for a comprehensive understanding human complex diseases like cancer are essential.
