Advances in the Development of Genetic Tools for the Genus Rhodococcus

Mitani, Yasuo; Nakashima, Naotoshi; Sallam, Khalid Ibrahim; Toriyabe, Tetsutya; Kondo, Kazuhiro; Tamura, Tomohide

doi:10.3209/saj.20.55

Cited by 8 publications

(3 citation statements)

References 71 publications

(75 reference statements)

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“…The production of specific proteins and functions in Rhodococcus spp. strains was achieved by developing several types of expression vectors based on cryptic (i.e., with unknown function) plasmids and transposable DNA elements tools for the insertion of cloned genes into chromosomes (Mitani et al 2006). In the field of bioconversions, Rhodococcus strains were engineered to improve the utilization of cellulose and hemicellulose degradation products such as cellobiose, xylose, l-arabinose, and levoglucosan (Fig.…”

Section: Heterologous Gene Expressionmentioning

confidence: 99%

Systems biology and metabolic engineering of Rhodococcus for bioconversion and biosynthesis processes

2021

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Rhodococcus spp. strains are widespread in diverse natural and anthropized environments thanks to their high metabolic versatility, biodegradation activities, and unique adaptation capacities to several stress conditions such as the presence of toxic compounds and environmental fluctuations. Additionally, the capability of Rhodococcus spp. strains to produce high value-added products has received considerable attention, mostly in relation to lipid accumulation. In relation with this, several works carried out omic studies and genome comparative analyses to investigate the genetic and genomic basis of these anabolic capacities, frequently in association with the bioconversion of renewable resources and low-cost substrates into triacylglycerols. This review is focused on these omic analyses and the genetic and metabolic approaches used to improve the biosynthetic and bioconversion performance of Rhodococcus. In particular, this review summarizes the works that applied heterologous expression of specific genes and adaptive laboratory evolution approaches to manipulate anabolic performance. Furthermore, recent molecular toolkits for targeted genome editing as well as genome-based metabolic models are described here as novel and promising strategies for genome-scaled rational design of Rhodococcus cells for efficient biosynthetic processes application.

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Section: Heterologous Gene Expressionmentioning

confidence: 99%

Systems biology and metabolic engineering of Rhodococcus for bioconversion and biosynthesis processes

2021

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“…The type strain of Rhodococcus erythropolis, JCM 3201, was originally isolated from soil as a degrader of aromatics (4). A lysozyme-sensitive mutant of JCM 3201, strain L88, has been used as an expression host of recombinant proteins that are difficult to express in Escherichia coli (5–9). JCM 3201 is also useful to the host for bioconversion of chemicals such as vitamin D 3 to hydroxyvitamin D 3 (10–12).…”

Section: Announcementmentioning

confidence: 99%

Genome Sequence of Rhodococcus erythropolis Type Strain JCM 3201

Yoshida

Kitagawa

Ishiya

et al. 2019

Microbiol Resour Announc

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“…Biocatalytic synthesis of acrylic monomers for the polymer industry using intact Rhodococcus cells − displaced earlier known chemical Cu-catalyzed synthesis, and bioconversion of lignocelluloses to renewable fuels and biobased chemicals using growing Rhodococcus cells is now promising for industrial applications . Owing to its unique metabolic potential, a wide range of potential practical applications, − and growing number of adapted genetic tools − rhodococci are now promising candidates for the “club” of microbial biocatalytic and biosynthetic platforms. To become a full-fledged member of the club, a set of characterized strains for different purposes is needed, along with a set of genetic parts for gene expression control characterized in these strains.…”

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confidence: 99%

A Set of Active Promoters with Different Activity Profiles for Superexpressing Rhodococcus Strain

et al. 2021

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Rhodococcus bacteria are a promising platform for biodegradation, biocatalysis, and biosynthesis, but the use of rhodococci is hampered by the insufficient number of both platform strains for expression and promoters that are functional and thoroughly studied in these strains. To expand the list of such strains and promoters, we studied the expression capability of the Rhodococcus rhodochrous M33 strain, and the functioning of a set of recombinant promoters in it. We showed that the strain supports superexpression of the target enzyme (nitrile hydratase) using alternative inexpensive feedingsacetate and ureawithout growth factor supplementation, thus being a suitable expression platform. The promoter set included P tuf (elongation factor Tu) and P sod (superoxide dismutase) from Corynebacterium glutamicum ATCC13032, P cpi (isocitrate lyase) from Rhodococcus erythropolis PR4, and P nh (nitrile hydratase) from R. rhodochrous M8. Activity levels, regulation possibilities, and growth-phase-dependent activity profiles of these promoters were studied in derivatives of the M33 strain. The activities of the promoters were significantly different (P cpi < P sod ≪ P tuf < P nh ), covering 10 3 -fold range, and the most active P nh and P tuf produced up to a 30−50% portion of target protein in soluble intracellular proteins. On the basis of the mRNA quantification and amount of target protein, the production level of P nh was positioned close to the theoretical upper limit of expression in a bacterial cell. A selection method for the laboratory evolution of such active promoters directly in Rhodococcus was also proposed. Concerning regulation, P tuf could not be regulated (2-fold change), while others were tunable (6-fold for P sod , 79-fold for P nh , and 44-fold for P cpi ). The promoters possessed four different activity profiles, including three with peak of activity at different growth phases and one with constant activity throughout the growth phases. P tuf and P cpi did not change their activity profile under different growth conditions, whereas the P sod and P nh profiles changed depending on the growth media. The results allow flexible construction of Rhodococcus strains using the studied promoters, and demonstrate a valuable approach for complex characterization of promoters intended for biotechnological strain construction.

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Advances in the Development of Genetic Tools for the Genus Rhodococcus

Cited by 8 publications

References 71 publications

Systems biology and metabolic engineering of Rhodococcus for bioconversion and biosynthesis processes

Systems biology and metabolic engineering of Rhodococcus for bioconversion and biosynthesis processes

Genome Sequence of Rhodococcus erythropolis Type Strain JCM 3201

A Set of Active Promoters with Different Activity Profiles for Superexpressing Rhodococcus Strain

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