Rpf protein, a kind of resuscitation promoting factor, was first found in the culture supernatant of Micrococcus luteus. It can resuscitate the growth of M. luteus in "viable but non-culture, VBNC" state and promote the growth of Gram-positive bacteria with high G + C content. This paper investigates the resuscitating activity of M. luteus ACCC 41016 Rpf protein, which was heterologously expressed in E. coli, to cells of M. luteus ACCC 41016 and Rhodococcus marinonascens HBUM200062 in VBNC state, and examines the effect on the cultivation of actinobacteria in soil. The results showed that the recombinant Rpf protein had resuscitation effect on M. luteus ACCC 41016 and R. marinonascens HBUM200062 in VBNC state. 83 strains of actinobacteria, which were distributed in 9 families and 12 genera, were isolated from the experimental group with recombinant Rpf protein in the culture medium. A total of 41 strains of bacteria, which were distributed in 8 families and 9 genera, were isolated from the control group without Rpf protein. The experimental group showed richer species diversity than the control group. Two rare actinobacteria, namely HBUM206391 and HBUM206404, were obtained in the experimental group supplemented with Rpf protein. Both may be potential new species of Actinomadura and Actinokineospora, indicating that the recombinant expression of M. luteus ACCC 41016 Rpf protein can effectively promote the isolation and culture of actinobacteria in soil.
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Article Synopsis
- KB1 is a petroleum-degrading bacterium from the Actinomycetota phylum that utilizes resuscitation-promoting factors (Rpf) to revive dormant cells and support growth.
- The study specifically investigates the Rpf2 protein in KB1, which has a complex structure with various domains, including DUF348, and examines how deletions of these domains affect its enzymatic and biological activities.
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