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首页> 外文期刊>BMC Biotechnology >A hyper-thermostable α-amylase from Pyrococcus furiosus accumulates in Nicotiana tabacum as functional aggregates
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A hyper-thermostable α-amylase from Pyrococcus furiosus accumulates in Nicotiana tabacum as functional aggregates

机译：激烈热球菌的超耐热α-淀粉酶以功能性聚集体的形式积累在烟草中

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BackgroundAlpha amylase hydrolyzes α-bonds of polysaccharides such as starch and produces malto-oligosaccharides. Its starch saccharification applications make it an essential enzyme in the textile, food and brewing industries. Commercially available α-amylase is mostly produced from Bacillus or Aspergillus . A hyper-thermostable and Ca 2++ independent α-amylase from Pyrococcus furiosus (PFA) expressed in E.coli forms insoluble inclusion bodies and thus is not feasible for industrial applications. ResultsWe expressed PFA in Nicotiana tabacum and found that plant-produced PFA forms functional aggregates with an accumulation level up to 3.4?g/kg FW (fresh weight) in field conditions. The aggregates are functional without requiring refolding and therefore have potential to be applied as homogenized plant tissue without extraction or purification. PFA can also be extracted from plant tissue upon dissolution in a mild reducing buffer containing SDS. Like the enzyme produced in P. furiosus and in E. coli , plant produced PFA preserves hyper-thermophilicity and hyper-thermostability and has a long shelf life when stored in lyophilized leaf tissue. With tobacco’s large biomass and high yield, hyper-thermostable α-amylase was produced at a scale of 42?kg per hectare. ConclusionsTobacco may be a suitable bioreactor for industrial production of active hyperthermostable alpha amylase.
机译：背景技术α淀粉酶水解诸如淀粉的多糖的α-键并产生麦芽寡糖。它的淀粉糖化应用使其成为纺织，食品和酿造行业必不可少的酶。市售的α-淀粉酶主要由芽孢杆菌或曲霉产生。在大肠杆菌中表达的来自激烈热球菌（PFA）的超耐高温且不依赖Ca 2 ++ 的α-淀粉酶形成不溶性包涵体，因此不适用于工业应用。结果我们在烟草中表达了PFA，发现在田间条件下，植物产生的PFA形成功能性聚集体，累积水平高达3.4？g / kg FW（鲜重）。聚集体是功能性的，不需要重新折叠，因此具有无需提取或纯化即可用作均质植物组织的潜力。当溶解在含有SDS的温和还原缓冲液中时，也可以从植物组织中提取PFA。像在狂热假单胞菌和大肠杆菌中产生的酶一样，植物产生的PFA保留超嗜热性和超恒温性，并且在冻干的叶片组织中储存时具有较长的保存期限。烟草具有巨大的生物量和高产，可产生每公顷42？kg的超耐热α-淀粉酶。结论烟草可能是工业化生产活性超热α淀粉酶的合适生物反应器。

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