The potential of Bacillus species isolated from Cinnamomum camphora for biofuel production.

BACKGROUND: Increasing concerns about climate change and global petroleum supply draw attention to the urgent need for the development of alternative methods to produce fuels. Consequently, the scientific community must devise novel ways to obtain fuels that are both sustainable and eco-friendly. Bacterial alkanes have numerous potential applications in the industry sector. One significant application is biofuel production, where bacterial alkanes can serve as a sustainable eco-friendly alternative to fossil fuels. This study represents the first report on the production of alkanes by endophytic bacteria. RESULTS: In this study, three Bacillus species, namely Bacillus atrophaeus Camph.1 (OR343176.1), Bacillus spizizenii Camph.2 (OR343177.1), and Bacillus aerophilus Camph.3 (OR343178.1), were isolated from the leaves of C. camphora. The isolates were then screened to determine their ability to produce alkanes in different culture media including nutrient broth (NB), Luria-Bertani (LB) broth, and tryptic soy broth (TSB). Depending on the bacterial isolate and the culture media used, different profiles of alkanes ranging from C8 to C31 were detected. CONCLUSIONS: The endophytic B. atrophaeus Camph.1 (OR343176.1), B. spizizenii Camph.2 (OR343177.1), and B. aerophilus Camph.3 (OR343178.1), associated with C. camphora leaves, represent new eco-friendly approaches for biofuel production, aiming towards a sustainable future. Further research is needed to optimize the fermentation process and scale up alkane production by these bacterial isolates.

Characterization of a novel lipase from Bacillus licheniformis NCU CS-5 for applications in detergent industry and biodegradation of 2,4-D butyl ester.

Enzymatic degradation has become the most promising approach to degrading organic ester compounds. In this study, Bacillus licheniformis NCU CS-5 was isolated from the spoilage of Cinnamomum camphora seed kernel, and its extracellular lipase was purified, with a specific activity of 192.98 U/mg. The lipase was found to be a trimeric protein as it showed a single band of 27 kDa in SDS-PAGE and 81 kDa in Native-PAGE. It was active in a wide range of temperatures (5-55 °C) and pH values (6.0-9.0), and the optimal temperature and pH value were 40 °C and 8.0, respectively. The enzyme was active in the presence of various organic solvents, metal ions, inhibitors and surfactants. Both crude and purified lipase retained more than 80% activity after 5 h in the presence of commercial detergents, suggesting its great application potential in detergent industry. The highest activity was found to be towards medium- and long-chain fatty acids (C6-C18). Peptide mass spectrometric analysis of the purified lipase showed similarity to the lipase family of B. licheniformis. Furthermore, it degraded more than 90% 2,4-D butyl ester to its hydrolysate 2,4-D within 24 h, indicating that the novel lipase may be applied to degrade organic ester pesticides.

Analysis of endophytic bacterial community diversity and metabolic correlation in Cinnamomum camphora.

The structure and diversity of microbial communities in the leaves of Cinnamomum camphora at different growth stages were studied by high-throughput sequencing. Moreover, the relationships between microbial communities and borneol content were analyzed in this paper. The results indicated that the community structure of endophytic bacteria in C. camphora exhibited temporal variations, with the microbial diversity presented as follows: T1 (low content period) > T3 (peak period) > T2 (small peak period). The population of endophytic bacteria and the ratio of primary metabolism in the leaves of C. camphora were T2 > T1 > T3, while the metabolic intensity of endophytic bacterial terpenoids and polyketides was T3 > T2 > T1, which had the same trend as borneol content in C. camphora. The metabolic ratio of terpenoids and polyketides in T3 was 7.44% higher than that in T1, while that in T2 was 4.10% higher than that in T1. The abundance and diversity of Clostridium_sensu_stricto_1, Ochrobactrum, Escherichia-Shigella, Pseudomonas, and Massilia significantly promoted the content of terpenoids in C. camphora. Together, those results provide the first evidence that borneol content and potential metabolic intensity in leaves of C. camphora greatly depend on microbial communities composition and diversity.

Pseudomonas hutmensis sp. nov., a New Fluorescent Member of Pseudomonas putida Group.

A rod-shaped, Gram-negative and aerobic bacterium, strain XWS2, was isolated from rhizosphere soil of a camphor tree in Hubei University of Chinese Medicine Huangjiahu Campus, Wuhan, China. Cells grew at 4-37 °C (optimum 28 °C), pH 5.0-9.0 (optimum 7.0) and with 0-5% NaCl (optimum 1%). Colonies growing on tryptone soybean agar are round, beige in color and approximately 2 mm in diameter after 24 h incubation at 28 °C. Pellicle formation during liquid culture and strong fluorescent pigment production on King's B medium are typical features of strain XWS2. The genome of XWS2 is 6,170,117 bp, containing 5682 predicted genes and 4770 genes are functionally annotated. Phylogenetic analysis based on 16S rRNA sequence showed that strain XWS2 formed an independent branch within the Pseudomonas putida group, with P. putida NBRC 14164T (99.86% similarity) and P. alkylphenolica KL28T (99.36% similarity) as the most closely related type strains. Genome sequence analysis based on average nucleotide identity and digital DNA-DNA hybridization are below the threshold values for species delineation. Phenotypic characteristics, physiological and biochemical tests also supported the strain represents a separate novel species within the Pseudomonas genus. The name Pseudomonas hutmensis sp. nov. is proposed, with type strain XWS2T (= CCTCC AB 2018189 T = KACC 19898T).

[Effects of nitrogen addition on red soil microbes in the Cinnamomum camphora plantation].

In order to investigate the effects of nitrogen addition on the red soil microbial communities in Cinnamomum camphora plantation, three treatments of nitrogen addition were designated as control (N0: 0 g x m(-2)), low nitrogen (N1: 5 g x m(-2)) and high nitrogen (N2 :15 g x m(-2)). Soil microbial numbers, microbial biomass carbon (C), biomass N and microbial community functional diversity were analyzed using the methods of plate counting, chloroform fumigation and BIOLOG system, respectively. The results showed that the numbers of bacteria in N1 and N2 were significantly higher than the control 1 month after nitrogen addition, but significantly lower than the control 13 months after nitrogen addition, and the number of fungi and actinomycetes were not significantly changed after nitrogen addition. The soil microbial biomass C, N increased with the increase of nitrogen at 1 month, but the soil microbial biomass C increased significantly 13 months after nitrogen addition when compared with 1 month after nitrogen addition. The soil microbial biomass N was lower 13 months after nitrogen addition when compared with 1 month after nitrogen addition, but the difference was not significant (P > 0.05). The variation of the carbon utilization efficiency of soil microbial communities was resulted from the nitrogen addition. The indices of Shannon index, Simpson index and McIntosh index were calculated to show the differences in nitrogen treatments and in times, which turned out to be insignificant.

[Volatile metabolites analysis and molecular identification of endophytic fungi bn12 from Cinnamomum camphora chvar. borneol].

OBJECTIVE: To identify endophytic fungi bn12 from Cinnamomum camphora chvar, borneol and analysis its volatile metabolites. METHOD: The endophytic fungi bn12 was identified by morphological observation. volatile metabolites of endophytic fungi bn12 was analyzed by gas chromatography/mass spectrography (GC-MS). RESULT: Volatile metabolites of endophytic fungi bn12 contain borneol and much indoles. The ITS sequence of endophytic fungi bnl2 is most similar to the ITS sequence of pleosporaceae fungus, particularly C. nisikadoi. CONCLUSION: Endophytic fungi bn12 is belong to pleosporaceae fungus. It has the ability of producing broneol.