The producing strain C4-6 was maintained on marine agar 2216 (Difco). A single colony of strain C4-6 was inoculated into a 500 mL K-1 flask containing 100 mL of marine broth 2216 (Difco) as a seed culture. The seed culture was incubated at 30 °C on a rotary shaker at 200 rpm for 2 days. Three mL of seed culture was inoculated into twenty-five 500 mL K-1 flasks each containing 100 mL of A11M production medium, which consists of glucose 0.2%, soluble starch 2.5%, yeast extract 0.5%, polypeptone (Wako Pure Chemical Industries, Ltd.) 0.5%, NZ-amine (Wako Pure Chemical Industries, Ltd.) 0.5%, CaCO3 0.3%, and Diaion HP-20 (Mitsubishi Chemical Co.) 1% in natural seawater (collected from Toyama Bay, Japan). The pH of the medium was adjusted to 7.0 before sterilization. The inoculated flasks were incubated at 30 °C on a rotary shaker at 200 rpm for 5 days.
Polypeptone
Manufactured by Fujifilm
Sourced in Japan, United States
Polypeptone is a culture medium component manufactured by Fujifilm. It is a complex mixture of polypeptides derived from the enzymatic hydrolysis of proteins. Polypeptone serves as a source of nitrogen, amino acids, and other growth factors for the cultivation of microorganisms in various laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Nz amine, by Fujifilm (2 mentions)
Diaion hp 20, by Mitsubishi (2 mentions)
Sp sepharose fast flow, by GE Healthcare (1 mentions)
Bca assay kit, by Beyotime (1 mentions)
Other analytical grade reagents, by Sinopharm (1 mentions)
Hipolypeptone, by Fujifilm (1 mentions)
Bordet gengou agar, by BD (1 mentions)
Yeast extract peptone dextrose, by BD (1 mentions)
Bacto yeast extract, by BD (1 mentions)
Yeast nitrogen base without amino acids, by BD (1 mentions)
Yeast extract, by BD (1 mentions)
Casamino acid, by BD (1 mentions)
7 protocols using polypeptone
1
Optimizing Microbial Marine Natural Product
2
Isolation and Characterization of Chryseobacterium cucumeris ZYF120413-7
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The Chryseobacterium cucumeris ZYF120413-7 strain was isolated from rice field soil according to the method of Yamaguchi and Yokoe (2000) (link) and stored in the Food and Microbial Technology Laboratory, School of Life Sciences, East China Normal University. SP-Sepharose FastFlow, Sephacryl S-100 HR pre-packed columns and the hollow fiber cross-flow filtration cartridges (UFP-5-C-3MA, MW 5KD) were purchased from GE Healthcare Life Sciences, United States. A Low molecular weight sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) marker was purchased from TaKaRa Bio Group, Japan. BCA assay kit was purchased from Beyotime Biotechnology, China. Polypeptone was purchased from Wako Pure Chemical Industries, Japan. SPI was obtained from Jinjing Biotechnology, China. Casein was obtained from Mercury Trading (Shanghai) Co., Ltd. Other analytical grade reagents were purchased from Sinopharm Group Chemical Reagent Co. Ltd. (Shanghai, China).
3
Isolation and Cultivation of Fusarium concentricum FKI-7550
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Fungal strain FKI-7550 was isolated from soil around the root of Cerasus × yedoensis collected in Tokushima, Japan. We have already reported that the fusaramin producing strain FKI-7550 was assigned to the genus and was designated as Fusarium concentricum (NITE-2944 [Sakai et al., 2019 (link)]). One loopful of strain Fusarium concentricum FKI-7550 grown on an LcA slant (0.1% glycerol, 0.08% KH2PO4, 0.02% K2HPO4, 0.02% MgSO4·7H2O, 0.02% KCl, 0.2% NaNO3, and 1.5% agar, pH 6.0) was inoculated into a 500 ml-Erlenmeyer flask containing 100 ml of a pre-seed culture medium [2% glucose, 0.2% yeast extract, 0.05% MgSO4·7H2O, 0.5% Polypeptone (FUJIFILM Wako Pure Chemical Co., Osaka, Japan), 0.1% KH2PO4, and 0.1% agar, pH 6.0] and incubated on a rotary shaker at 27°C for 4 days. Fifty-hundred ml of seed culture was prepared by the pre-seed culture medium. Twenty-five ml of the seed culture was inoculated into each of 200 Ulpack 47 culture bags (Hokken Co., Ltd., Tochigi, Japan) containing a production medium (500 g of water-sodden rice). Static fermentation was continued at 27°C for 12 days.
4
Cultivation of Marine Bacterium T35-5
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In a similar manner as described in [22 (link)], the strain T35-5 was maintained on Marine Agar 2216 (Difco). A loopful of the strain T35-5 was inoculated into a 500 mL K-1 flask containing 100 mL of Marine Broth 2216 (Difco) as a seed culture. The seed culture was incubated at 30 °C on a rotary shaker at 200 rpm for 2 days. Three mL each of the seed culture were inoculated into 500 mL K-1 flasks containing 100 mL of A11M production medium, which consists of 0.2% glucose, 2.5% soluble starch, 0.5% yeast extract, 0.5% polypeptone (Wako Pure Chemical Industries, Ltd.), 0.5% NZ-amine (Wako Pure Chemical Industries, Ltd.), 0.3% CaCO3, and 1% Diaion HP-20 (Mitsubishi Chemical Co.) in natural seawater (collected from Toyama Bay, Japan). The pH value of the medium was adjusted to 7.0 before sterilization. The inoculated flasks were incubated at 30 °C for 5 days, with rotational shaking at 200 rpm.
5
Cultivation and Mutant Generation of B. pertussis
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B. pertussis strain Tohama I was maintained in the laboratory. A dnt-deficient mutant of B. pertussis Tohama I was generated according to methods described previously (45 (link)). B. pertussis was grown in Stainer-Scholte (SS) medium or on Bordet-Gengou agar (Becton, Dickinson, Franklin Lakes, NJ, USA) containing 0.4% (wt/vol) polypeptone or Hipolypeptone (Wako Pure Chemical Industries, Ltd., Japan), 0.8% glycerol, 20% defibrinated horse blood, and 10 μg/ml ceftibuten (BG plate). The culture supernatants of B. pertussis were harvested by centrifugation at 6,800 × g for 5 min and filtered through a 0.22-μm-pore-size filter.
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6
Yeast Mannoprotein Deletion Mutants
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Of 36 mannoprotein gene-deletion mutants of the budding yeast, S. cerevisiae [3 (link)], we studied 32 mutants (Table S1 ) that were straightforward to observe as single cells under a microscope. These 32 mutants were isogenic derivatives of BY4741 (MATa his3 leu2 met15 ura3) and were purchased from EUROSCARF (Oberursel, Germany). Cells with mutations in the other four mannoprotein genes (aga1∆, flo5∆, flo9∆, and dan4∆) were not studied due to heavy cell aggregation. Mutant and wild-type (WT) strains were cultured under optimal growth conditions at 25 °C in nutrient-rich yeast extract peptone dextrose (YPD) medium containing 1% (w/v) Bacto yeast extract (BD Biosciences, San Jose, CA, USA), 2% (w/v) polypeptone (Wako Chemicals, Richmond, VA, USA), and 2% (w/v) dextrose, as described previously [14 (link)]. A WT diploid strain (BY4743) and the homozygous gene deletion mutants in the BY4743 background used for Western blotting were purchased from EUROSCARF (Oberursel, Germany).
7
Yeast Cell Culture and Susceptibility Assays
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Yeast cells were routinely cultured overnight in yeast extract peptone dextrose (YPD) medium (2% glucose, 2% peptone, and 1% yeast extract) at 220 rpm and 24℃ to an absorbance at 600 nm (A 600 ) of 0.8-1. The culture was refreshed to an A 600 of 0.2 using YPD, incubated for 2 hour and divided into two parts. One part was allowed to continue growing under the same conditions (nontreated culture), and the other was supplemented with PA (a gift from Fachuang Lu, Ruili Gao, Jeff Piotrowski, and John Ralph at The University of Wisconsin) to a final concentration of 100 µg/mL. The cells were collected and subsequently processed according to the experimental approach.
For metachromatic interaction and PA susceptibility experiments, S. cerevisiae and Candida albicans were first cultured on YPD agar (1% yeast extract [BD Biosciences, San Jose, CA], 2% polypeptone [Fujifilm Wako Pure Chemical Corporation, Osaka, Japan], 2% glucose [Fujifilm Wako Pure Chemical Corporation, Osaka, Japan] and 2% agar [Fujifilm Wako Pure Chemical Corporation, Osaka, Japan]). Single colonies were inoculated into rich medium YPD or synthetic dextrose (SD; 0.67% yeast nitrogen base without amino acids [BD Biosciences] and 2% glucose) growth medium supplemented appropriately and incubated at 30 or 25℃ with shaking at 200 rpm. For Ura or Leu selection, 0.5% casamino acids (BD Biosciences) was added to SD medium.
For metachromatic interaction and PA susceptibility experiments, S. cerevisiae and Candida albicans were first cultured on YPD agar (1% yeast extract [BD Biosciences, San Jose, CA], 2% polypeptone [Fujifilm Wako Pure Chemical Corporation, Osaka, Japan], 2% glucose [Fujifilm Wako Pure Chemical Corporation, Osaka, Japan] and 2% agar [Fujifilm Wako Pure Chemical Corporation, Osaka, Japan]). Single colonies were inoculated into rich medium YPD or synthetic dextrose (SD; 0.67% yeast nitrogen base without amino acids [BD Biosciences] and 2% glucose) growth medium supplemented appropriately and incubated at 30 or 25℃ with shaking at 200 rpm. For Ura or Leu selection, 0.5% casamino acids (BD Biosciences) was added to SD medium.
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