Generation of incednine derivatives by mutasynthesis

Mutational Biosynthesis of Hitachimycin Analogs Controlled by the β-Amino Acid–Selective Adenylation Enzyme HitB

“…17 It is also carefully acknowledged that downstream PKS modules often reject unnatural types of polyketide chains in engineered polyketide biosynthesis. 18 Similarly, (S)-3-cylcohexyl-3APA (19) and 3AHA (20) had relatively low K M values of 130 μM and 140 μM, respectively. However, those K M values (>100 μM) may be too high to be recognized by HitB for efficient mutasynthesis.…”

“…A similar observation has been also reported for the mutasynthesis of incednine analogs, in which 3-aminopentanoic acid was successfully substituted for the starter unit moiety and led to the production of an incednine analog, while feeding with β-alanine led to the production of only a trace amount of the product. 19 The adenylation enzyme IdnL1, which is a homologue of HitB that participates in incednine biosynthesis, recognizes (S)-3-aminobutyric acid as a natural substrate and can also bind 3aminopentanoic acid but not β-alanine much. 20 Crystal Structural Analysis of the Adenylation Enzyme HitB.…”

“…The ΔhitA strain of S. scabrisporus JCM 11712 was cultured in 0.5 × TSB medium (0.85% tryptone, 0.15% soytone, 0.125% D-glucose, 0.25% NaCl, and 0.125% KH 2 PO 4 ) at 28 °C with shaking at 200 rpm for 3 days for the seed culture and in 0.3 × BPS medium (0.9% oat meal, 0.15% malt extract, 0.09% yeast extract, 0.012% MgSO 4 •H 2 O, 0.15% CaCO 3 , 0.03% NaCl, 0.9% soluble starch, 1.1% MOPS, and 1% D-glucose) at 28 °C with shaking at 200 rpm for 7 days for the main culture as previously described, except for feeding with the (S)-β-Phe analogs. 13 10), (S)-β-3,5-dimethyl-Phe (25), and (S)-3-cyclohexyl-3APA (19) were purchased from Chemspace (https://chem-space.com). (S)-β-m-Br-Phe ( 6), (S)-β-m-CN-Phe ( 17), (S)-β-m-OCH 3 -Phe (12), and (R)-3-aminohex-5-enoic acid (29) were purchased from Watanabe Chemical Co., Ltd. (Osaka, Japan).…”

Postępy w rozwoju narzędzi biologii syntetycznej do bioprodukcji analogów produktów naturalnych

“…W konsekwencji uzyskano 28-metyloincedninę poprzez wprowadzenie kwasu 3-aminopentanowego do hodowli szczepu, w którym doszło do przerwania genu glutaminianu 2,3-aminomutazy, który odpowiadał za dostarczanie kwasu. 28-metyloincednina wykazała także działanie hamujące funkcję antyapoptyczną onkoproteiny, dowodząc przydatności mutasyntezy w produkcji poliketydów zawierających β-aminokwasy [36]. Podobnie cytochalazyna, która hamuje polimeryzację aktyny tworzącej mikrofilamenty cytoszkieletu oraz transport monosacharydów przez błonę komórkową i jej pochodne bez konieczności stosowania strategii ochronnych, zostały uzyskane z mutanta Magnaporthe grisea.…”