Efficient and scalable synthesis of 1,5-diamino-2-hydroxy-pentane from l-lysine via cascade catalysis using engineered Escherichia coli

Abstract

Background 1,5-Diamino-2-hydroxy-pentane (2-OH-PDA), as a new type of aliphatic amino alcohol, has potential applications in the pharmaceutical, chemical, and materials industries. Currently, 2-OH-PDA production has only been realized via pure enzyme catalysis from lysine hydroxylation and decarboxylation, which faces great challenges for scale-up production. However, the use of a cell factory is very promising for the production of 2-OH-PDA for industrial applications, but the substrate transport rate, appropriate catalytic environment (pH, temperature, ions) and separation method restrict its efficient synthesis. Here, a strategy was developed to produce 2-OH-PDA via an efficient, green and sustainable biosynthetic method on an industrial scale. Results In this study, an approach was created for efficient 2-OH-PDA production from l-lysine using engineered E. coli BL21 (DE3) cell catalysis by a two-stage hydroxylation and decarboxylation process. In the hydroxylation stage, strain B14 coexpressing l-lysine 3-hydroxylase K3H and the lysine transporter CadB-argT enhanced the biosynthesis of (2S,3S)-3-hydroxylysine (hydroxylysine) compared with strain B1 overexpressing K3H. The titre of hydroxylysine synthesized by B14 was 2.1 times higher than that synthesized by B1. Then, in the decarboxylation stage, CadA showed the highest hydroxylysine activity among the four decarboxylases investigated. Based on the results from three feeding strategies, l-lysine was employed to produce 110.5 g/L hydroxylysine, which was subsequently decarboxylated to generate a 2-OH-PDA titre of 80.5 g/L with 62.6% molar yield in a 5-L fermenter. In addition, 2-OH-PDA with 95.6% purity was obtained by solid-phase extraction. Thus, the proposed two-stage whole-cell biocatalysis approach is a green and effective method for producing 2-OH-PDA on an industrial scale. Conclusions The whole-cell catalytic system showed a sufficiently high capability to convert lysine into 2-OH-PDA. Furthermore, the high titre of 2-OH-PDA is conducive to separation and possesses the prospect of industrial scale production by whole-cell catalysis.

Engineering an Artificial Pathway to Improve the Bioconversion of Lysine into Chiral Amino Alcohol 2-Hydroxycadaverine Using a Semi-Rational Design

“…Recently, a two-stage biocatalytic approach for the direct production of 2HyC from L-lysine via whole-cell biotransformation was first established by Li et al [27]. In the decarboxylation step, decarboxylase CadA can decarboxylate not only 3-hydroxylysine but also lysine [27]. Therefore, 2HyC can only be synthesized using a two-stage biocatalytic approach in this research [27].…”

“…However, pure enzyme catalysts are limited in their application due to their cumbersome purification steps and high cost [33]. Recently, a two-stage biocatalytic approach for the direct production of 2HyC from L-lysine via whole-cell biotransformation was first established by Li et al [27]. In the decarboxylation step, decarboxylase CadA can decarboxylate not only 3-hydroxylysine but also lysine [27].…”

“…As shown in Figure 1B, bifunctional compounds of chiral AAs are widely applied in many important fields of medicine, fine chemicals, materials, and asymmetric catalysis [26]. AAs are the precursor of polypeptide drugs such as Octreotide, Ziconotide, and Alamethicin F-30, nucleotide drugs such as Glycopeptidolipid, quinolone drugs such as Levofloxacin and Pazufloxacin, and radioactive drugs such as Lopamidol [27]. In addition, AAs have low toxicity and low viscosity, inhibit corrosion, absorb carbon dioxide, and cure at room temperature.…”

“…First, lysine was hydroxylated to generate 3-hydroxylysine by lysine hydroxylase. Subsequently, 3-hydroxylysine was decarboxylated by decarboxylase to form 2HyC [21,27]. Baud et al reported the direct synthetic route of 2HyC from L-lysine via enzymatic reactions using purified lysine hydroxylase KDO1 and purified lysine decarboxylase [21].…”

“…However, pure enzyme catalysts are limited in their application due to their cumbersome purification steps and high cost [33]. Li et al first established a two-stage biocatalytic approach for the direct production of 2HyC from L-lysine via whole-cell biotransformation [27].…”