A new trans‐4‐hydroxy‐l‐proline (trans‐Hyp) producing Bacillus cereus HBL‐AI, was isolated from the air, which was screened just using l‐proline as carbon and energy sources. This strain exhibited 73·4% bioconversion rate from initial l‐proline (3 g l−1) to trans‐Hyp. By sequencing the genome of this bacterium, 6244 coding sequences were obtained. Genome annotation analysis and functional expression were used to identify the proline‐4‐hydroxylase (BP4H) in HBL‐AI. This enzyme belonged to a family of 2‐oxoglutarate‐related dioxygenases, which required 2‐oxoglutarate and O2 as co‐substrates for the reaction. Homologous modelling indicated that the enzyme had two monomers and contained conserved motifs, which included a distorted ‘jelly roll’ β strand core and the residues (HXDXnH and RXS). The engineering Escherichia coli 3 Δ W3110/pTrc99a‐proba‐bp4h was constructed using BP4H, which transformed glucose to trans‐Hyp in one step with high concentration of 46·2 g l−1. This strategy provides a green and efficient method for synthesis of trans‐Hyp and thus has a great potential in industrial application.

中文翻译：

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蜡样芽孢杆菌 HBL-AI 菌株的分离及其在反式 4-羟基-L-脯氨酸生产中的应用

从空气中分离出一种新的生产反式-4-羟基-l-脯氨酸（trans-Hyp）的蜡状芽孢杆菌HBL-AI，仅使用l-脯氨酸作为碳和能源进行筛选。从最初的 l-脯氨酸 (3 gl-1) 到 trans-Hyp，该菌株表现出 73·4% 的生物转化率。通过对该细菌的基因组进行测序，获得了6244个编码序列。使用基因组注释分析和功能表达来鉴定 HBL-AI 中的脯氨酸 4-羟化酶 (BP4H)。该酶属于 2-氧戊二酸相关双加氧酶家族，需要 2-氧戊二酸和 O2 作为反应的共底物。同源模型表明该酶具有两个单体并包含保守基序，其中包括扭曲的“果冻卷”β 链核心和残基（HXDXnH 和 RXS）。使用 BP4H 构建工程大肠杆菌 3 Δ W3110/pTrc99a-proba-bp4h，将葡萄糖一步转化为 trans-Hyp，浓度为 46·2 gl-1。该策略为反式Hyp的合成提供了一种绿色、高效的方法，具有巨大的工业应用潜力。