High Throughput Cultivation for Isolation of Novel Marine Microorganisms

Toledo, Gerardo; Green, Wayne; González, Ricardo Castro; Christoffersen, L.; Podar, Mircea; Chang, Hwai W.; Hemscheidt, Thomas; Trapido-Rosenthal, Henry G.; Short, Jay M.; Bidigare, Robert R.; Mathur, Eric J.

doi:10.5670/oceanog.2006.75

Cited by 19 publications

(7 citation statements)

References 18 publications

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“…Furthermore, the isolates were de-replicated via Fourier transform infrared (FT/IR) spectroscopy. This study showed that 42% of the isolated strains obtained by using the microcapsule-based cultivation were novel (<98% 16S rRNA gene identity) as compared to 7% using the traditional agar plate isolation technique ( Toledo et al, 2012 ). Furthermore, the use of FT/IR spectroscopy to de-replicate isolates before identification reduced the time and cost investment to obtain novel isolates.…”

Section: Cultivation Techniques Used For Isolation Of Sponge-associated Bacteriamentioning

confidence: 96%

“…Single encapsulated cells that are able to grow under these conditions form microcolonies within the microcapsules (Zengler et al, 2002(Zengler et al, , 2005. Gerardo Toledo et al (2012) applied this method to cultivate bacteria from the sponge Mycale armata. Furthermore, the isolates were de-replicated via Fourier transform infrared (FT/IR) spectroscopy.…”

Section: Other Solid Substratesmentioning

confidence: 99%

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Bacteria Cultivated From Sponges and Bacteria Not Yet Cultivated From Sponges—A Review

et al. 2021

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The application of high-throughput microbial community profiling as well as “omics” approaches unveiled high diversity and host-specificity of bacteria associated with marine sponges, which are renowned for their wide range of bioactive natural products. However, exploration and exploitation of bioactive compounds from sponge-associated bacteria have been limited because the majority of the bacteria remains recalcitrant to cultivation. In this review, we (i) discuss recent/novel cultivation techniques that have been used to isolate sponge-associated bacteria, (ii) provide an overview of bacteria isolated from sponges until 2017 and the associated culture conditions and identify the bacteria not yet cultured from sponges, and (iii) outline promising cultivation strategies for cultivating the uncultivated majority of bacteria from sponges in the future. Despite intensive cultivation attempts, the diversity of bacteria obtained through cultivation remains much lower than that seen through cultivation-independent methods, which is particularly noticeable for those taxa that were previously marked as “sponge-specific” and “sponge-enriched.” This poses an urgent need for more efficient cultivation methods. Refining cultivation media and conditions based on information obtained from metagenomic datasets and cultivation under simulated natural conditions are the most promising strategies to isolate the most wanted sponge-associated bacteria.

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Section: Cultivation Techniques Used For Isolation Of Sponge-associated Bacteriamentioning

confidence: 96%

Section: Other Solid Substratesmentioning

confidence: 99%

Bacteria Cultivated From Sponges and Bacteria Not Yet Cultivated From Sponges—A Review

et al. 2021

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“…Marine bacteria have been attracting more attention as a resource for new bioactive compounds recently, as the soil-derived compounds have yielded high numbers of the previously described metabolites (Toledo et al 2006;Bhatnagar and Kim 2010;Trincone 2011). The present study was conducted to optimize a marine bacterium chitinase production and examine its properties.…”

Section: Resultsmentioning

confidence: 99%

A novel halo-alkali-tolerant and thermo-tolerant chitinase from Pseudoalteromonas sp. DC14 isolated from the Caspian Sea

Makhdoumi

Dehghani-Joybari

Mashreghi

et al. 2015

Int. J. Environ. Sci. Technol.

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A novel halo-alkali and thermo-tolerant chitinase was obtained from an isolated strain found in the Caspian Sea. The effects of media composition and various fermentation conditions for the optimization of chitinase production were studied one factor at a time and by response surface methodology. The novel strain, which is designated as strain DC14 and phylogenetically related to the genus Pseudoalteromonas, produced chitinase after 72 h under the following optimal conditions: glucose 1 % (w/v), ammonium sulphate 0.2 % (w/v), chitin 1.07 % (w/ v), pH 8, NaCl 10 % (w/v), inoculums size 2.5 % (v/v), temperature 30°C, CaCl 2 3 mM and MgCl 2 4 mM. Using the statistical optimization method, chitinase production was found to increase from 2.30 to 21.90 U/dl. The enzyme showed maximum activity at 40°C, pH 9 and 10 % NaCl. It was stable in a wide range of temperature from 15 to 65°C, pH from pH 7 to 11 and NaCl concentration from 0 to 15 % (w/v). The molecular weight of the enzyme was estimated by SDS-PAGE to be about 65 kDa. With regard to the halo-alkali and thermo-stable properties of this enzyme, it has potential industrial activity.

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“…The marine environments characterized by high salinity, high pressure, and low temperature hold the promise of producing new natural products structurally or bioactively distinct with those from terrestrial environments. Various Bacillus species have been isolated from marine sources with different isolation methods [ 14 , 15 , 16 ], and new nonribosomal peptides with varied bioactivities have been discovered. Gageopeptides that were discovered from Bacillus subtilis collected from Gageocho reef, Republic of Korea were linear lipopeptides showing potent antifungal and antibacterial activities [ 17 ].…”

Section: Introductionmentioning

confidence: 99%

Bacillibactin and Bacillomycin Analogues with Cytotoxicities against Human Cancer Cell Lines from Marine Bacillus sp. PKU-MA00093 and PKU-MA00092

et al. 2018

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Nonribosomal peptides from marine Bacillus strains have received considerable attention for their complex structures and potent bioactivities. In this study, we carried out PCR-based genome mining for potential nonribosomal peptides producers from our marine bacterial library. Twenty-one “positive” strains were screened out from 180 marine bacterial strains, and subsequent small-scale fermentation, HPLC and phylogenetic analysis afforded Bacillus sp. PKU-MA00092 and PKU-MA00093 as two candidates for large-scale fermentation and isolation. Ten nonribosomal peptides, including four bacillibactin analogues (1–4) and six bacillomycin D analogues (5–10) were discovered from Bacillus sp. PKU-MA00093 and PKU-MA00092, respectively. Compounds 1 and 2 are two new compounds and the 1H NMR and 13C NMR data of compounds 7 and 9 is first provided. All compounds 1–10 were assayed for their cytotoxicities against human cancer cell lines HepG2 and MCF7, and the bacillomycin D analogues 7–10 showed moderate cytotoxicities with IC50 values from 2.9 ± 0.1 to 8.2 ± 0.2 µM. The discovery of 5–10 with different fatty acid moieties gave us the opportunity to reveal the structure-activity relationships of bacillomycin analogues against these human cancer cell lines. These results enrich the structural diversity and bioactivity properties of nonribosomal peptides from marine Bacillus strains.

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High Throughput Cultivation for Isolation of Novel Marine Microorganisms

Cited by 19 publications

References 18 publications

Bacteria Cultivated From Sponges and Bacteria Not Yet Cultivated From Sponges—A Review

Bacteria Cultivated From Sponges and Bacteria Not Yet Cultivated From Sponges—A Review

A novel halo-alkali-tolerant and thermo-tolerant chitinase from Pseudoalteromonas sp. DC14 isolated from the Caspian Sea

Bacillibactin and Bacillomycin Analogues with Cytotoxicities against Human Cancer Cell Lines from Marine Bacillus sp. PKU-MA00093 and PKU-MA00092

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