Tryptone

Manufactured by Lab M

Sourced in United Kingdom

Tryptone is a peptic digest of casein, a protein derived from milk. It is a commonly used nutrient source for the cultivation of various microorganisms in microbiology and biotechnology laboratories. Tryptone provides a complex mixture of amino acids, peptides, and other growth-promoting compounds that support the growth and metabolic activities of a wide range of bacterial and fungal species.

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Close spelling variants of this product

Tryptone Soya Agar (TSA) (6 citations) Tryptone Soy Broth (4 citations) Tryptone Soy Broth (TSB; (3 citations) Tryptone Soy Agar (TSA, (2 citations) Tryptone soya broth (2 citations)

9 protocols using tryptone

Bacterial Growth Media and Antibiotics

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The bacterial strains and plasmids used in this study are described in Table 1 and used oligonucleotides in Table S1. Bacteria were grown in a complete LB medium containing 10 g/L tryptone (LabM, Heywood, UK), 5 g/L yeast extract (LabM, Heywood, UK) and 5 g/L NaCl, or in media based on M9 buffer [38 ]. The M9-0.2CAA medium contained M9 buffer [38 ], 2.5 mL/L of microelements [39 (link)], 2 g/L of glucose and 2 g/L of CAA (casamino acid; Difco, Detroit, MI, USA) and 0.01 g/L tryptophan. M9-0.2CAA was amended with 0.4 mg/mL poly-L-lysine (1000–5000 Da; Sigma-Aldrich, St. Louis, MO, USA); or biopolymers: with 10 g/L tryptone (LabM, Heywood, UK), with 0.4 g/L the sodium salt of carboxyl methylcellulose (Sigma-Aldrich, St. Louis, MO, USA) or PGA (poly-galacturonic acid). All used growth media are summarised in Table S2.
P. putida was incubated at 30 °C, E. coli at 37 °C. Solid media contained 1.5% agar (Difco, Detroit, MI, USA). Antibiotics were added at the following concentrations: 10 µg gentamicin mL⁻¹, 100 µg ampicillin mL⁻¹, 50 µg kanamycin mL⁻¹, 1.5 mg benzylpenicillin mL⁻¹, 200 µg streptomycin mL⁻¹.

Puhm M., Hendrikson J., Kivisaar M, & Teras R. (2022). Pseudomonas putida Biofilm Depends on the vWFa-Domain of LapA in Peptides-Containing Growth Medium. International Journal of Molecular Sciences, 23(11), 5898.

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Kombucha SCOBY Cultivation Protocol

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Kombucha SCOBY (symbiotic colony of bacteria and yeast) was obtained from Sri Dhanvanthiri Probiotics Ltd., Kodaikanal, India (True Brew Kombucha tea). Sodium chloride, sodium molybdate, potassium chloride, di sodium hydrogen phosphate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, calcium carbonate, calcium chloride, magnesium sulfate, disodium hydrogen phosphate, citric acid, bromothymol blue and oxidase test disks (product no: 70439) were purchased from Merck (Darmstadt, Germany). Acetic acid and agar were purchased from Fisher Scientific (Loughborough, UK). Glucose and casein amino acids were purchased from VWR International (Leuven, Belgium). Tryptone, peptone and yeast extract were from Lab M Limited (Heywood, UK). Ethanol was from Altia Oyj (Helsinki, Finland). Cycloheximide (Product no: C7698), and cellulase from Trichoderma reesei ATCC 26921 (product no: C2730) was purchased from Sigma-Aldrich (St. Louis, MO, USA). GeneJET Genomic DNA Purification Kit was purchased from Thermo Scientific (Waltham, MA, USA). Crude glycerol was generously provided by Perstorp AB (Malmo, Sweden).

Cannazza P., Rissanen A.J., Guizelini D., Losoi P., Sarlin E., Romano D., Santala V, & Mangayil R. (2021). Characterization of Komagataeibacter Isolate Reveals New Prospects in Waste Stream Valorization for Bacterial Cellulose Production. Microorganisms, 9(11), 2230.

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Synthesis and Characterization of Ag-Doped Nanoparticles

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Sodium chloride (NaCl, ≥99%, Taufkirchen, Germany), sodium hydroxide (NaOH, Prague, Czech Republic), silver nitrate (AgNO₃, >99.8%, Gillingham, UK), 2′, 7′ Dichlorofluorescin diacetate (C₂₄H₁₆Cl₂O₇, ≥98%, Rehovot, Israel), ethylene-co-vinyl acetate polymer (EVA, 40 wt.%, Taufkirchen, Germany), dichloromethane (CH₂Cl₂, Taufkirchen, Germany), poly(vinyl pyrrolidone) (PVP, Mw 1,300,000 by LS, St. Louis, MO, USA) and phosphate buffered saline (PBS, pH 7.4, Taufkirchen, Germany) were purchased from Sigma Aldrich and used as received. Iron (III) nitrate nonahydrate (Fe(NO₃)₃∙9H₂O, ≥98%, Darmstadt, Germany), hexane (C₆H₁₄, ≥97.0%, Rosh-Ha’ayin, Israel), and Nutrient agar (NA, Darmstadt, Germany) were purchased from Merck. Tryptone, yeast extract, meat extract, and agar were obtained from LabM (Heywood, UK). All chemicals were purchased as analytical reagent (AR) grade and used without further purification. High purity deionized (DI) water obtained from a Milli-Q^® system (Merck Millipore, Darmstadt, Germany) was used for all the tests.

Vihodceva S., Šutka A., Otsus M., Vija H., Grase L., Kahru A, & Kasemets K. (2022). Visible-Light Active Flexible and Durable Photocatalytic Antibacterial Ethylene-co-vinyl Acetate—Ag/AgCl/α-Fe2O3 Composite Coating. Nanomaterials, 12(12), 1984.

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Antibiotic Susceptibility of Pseudomonas aeruginosa

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Luria-Bertani (LB) agar, LB broth and tryptone were purchased from Lab M Limited (Lancashire, UK). Mueller Hinton agar, Mueller Hinton broth, and tryptone soya agar and tryptone soya broth were obtained from Oxoid (Hampshire, UK). Other chemicals were of pharmaceutical grade. Bacterial strains of Pseudomonas aeruginosa PAO1 were kindly donated by the Department of Microbiology, Faculty of Pharmacy, Mansoura University, Egypt.

Mostafa I., Abbas H.A., Ashour M.L., Yasri A., El-Shazly A.M., Wink M, & Sobeh M. (2020). Polyphenols from Salix tetrasperma Impair Virulence and Inhibit Quorum Sensing of Pseudomonas aeruginosa. Molecules, 25(6), 1341.

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Oxidative Stress Assay Protocol

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Iron (III) nitrate nonahydrate (Fe(NO₃)₃∙9H₂O, ≥98%, Steinheim, Germany), sodium chloride (NaCl, ≥99%, Steinheim, Germany), trisodium citrate (C₆H₅Na₃O_7, Steinheim, Germany), 2′, 7′ Dichlorofluorescin diacetate (C₂₄H₁₆Cl₂O₇, ≥98%, Jerusalemci, Israel) and hydrogen peroxide solution (H₂O₂, 30%, Steinheim, Germany) were purchased from Sigma Aldrich and used as received. High purity deionized (DI) water obtained from a Milli-Q^® system (Merck Millipore, Darmstadt, Germany) was used for all the tests. Mueller–Hinton agar (MHA) was purchased from Sigma Aldrich (Bangalore, India). Tryptone, yeast extract, and agar were obtained from LabM (Lancashire, UK).

Vihodceva S., Šutka A., Sihtmäe M., Rosenberg M., Otsus M., Kurvet I., Smits K., Bikse L., Kahru A, & Kasemets K. (2021). Antibacterial Activity of Positively and Negatively Charged Hematite (α-Fe2O3) Nanoparticles to Escherichia coli, Staphylococcus aureus and Vibrio fischeri. Nanomaterials, 11(3), 652.

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Bacterial and Yeast Cultivation Protocols

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The strains used in this study are shown in Table 2. Escherichia coli DH5α (New-England-Biolabs (NEB)) was used for cloning and plasmid amplification. Bacteria were propagated in low-salt lysogeny broth (LS-LB) medium, consisting of 0.5% (w/v) sodium chloride (Merck), 0.5% (w/v) yeast extract (Lab M) and 1.0% (w/v) tryptone (Lab M) with or without 1.5% agar (BD) and with the required antibiotics. The Pichia pastoris (syn. Komagataella phaffii)
NRRL-Y-11430 strain (kindly obtained from Prof. Nico Callewaert (VIB, Ghent University)) was used for recombinant protein production. Yeast strains were plated on yeast extract peptone dextrose (YPD) plates (1% (w/v) yeast extract, 2% (w/v) peptone (BD), 2% (w/v) dextrose (Merck), 1.5% agar (Difco)) with the required antibiotics. All antibiotics were purchased from Thermo Fischer Scientific with the exception of carbenicillin, which was obtained from Gold Biotechnology. For recombinant protein expression, strains were grown in buffered glycerol-complex medium (BMGY) and induction was performed in buffered methanol-complex medium (BMMY). Both BMGY and BMMY consist of 1% (w/v) yeast extract, 2% (w/v) peptone (BD), 100 mM phosphate buffer (Sigma-Aldrich) at pH 6.0 and 1.34% (w/v) yeast nitrogen base (YNB, Formedium) with 1% glycerol (v/v Sigma) or 1% methanol (v/v, VWR) as sole carbon source respectively.

De Waele S., Vandenberghe I., Laukens B., Planckaert S., Verweire S., Van Bogaert I.N.A., Soetaert W., Devreese B, & Ciesielska K. (2018). Optimized expression of the Starmerella bombicola lactone esterase in Pichia pastoris through temperature adaptation, codon-optimization and co-expression with HAC1. Protein expression and purification, 143.

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Analytical-Grade Chemical Usage in Bio-Assays

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All the purchased chemicals were at least of analytical grade. Dulbecco’s phosphate-buffered saline (DPBS, Biognost), Alamar Blue (AppliChem), CuSO₄ (Alfa Aesar), 2′,7′-dichlorodihydrofluorescein diacetate (H₂DCF-DA, Life Technologies), phosphate buffered saline (PBS pH = 7.2, Biognost), tryptone (LabM), yeast extract (LabM), agar (LabM) and NaCl (Sigma-Aldrich) were used.

Kubo A.L., Vasiliev G., Vija H., Krishtal J., Tõugu V., Visnapuu M., Kisand V., Kahru A, & Bondarenko O.M. (2020). Surface carboxylation or PEGylation decreases CuO nanoparticles’ cytotoxicity to human cells in vitro without compromising their antibacterial properties. Archives of Toxicology, 94(5), 1561-1573.

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Cytotoxicity Assessment of Silver Nanoparticles

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All the purchased chemicals were at least of analytical grade. Media components: yeast extract, tryptone, agar were from Lab M (Lancashire, UK) and peptone was from BD. Phosphate buffered saline (PBS), Dulbecco's Modified Eagle's Medium (DMEM) with high glucose content (4.5 g/L), Newborn calf serum (NBCS) and the mixture of penicillin and streptomycin (10 000 U/mL or 10 000 µg/mL, respectively) were purchased from Life Technologies. Neutral red (NR) was from Applichem. AgNO₃ was purchased as 0.1 M solution from Fluka, 2,7- dichlorodihydrofluorescein diacetate from Invitrogen. Ultrapure water (UP water, pH 5.6±0.1) from MilliQ equipment (18 MΩ) was used throughout the study.

Ivask A., Kurvet I., Kasemets K., Blinova I., Aruoja V., Suppi S., Vija H., Käkinen A., Titma T., Heinlaan M., Visnapuu M., Koller D., Kisand V, & Kahru A. (2014). Size-Dependent Toxicity of Silver Nanoparticles to Bacteria, Yeast, Algae, Crustaceans and Mammalian Cells In Vitro. PLoS ONE, 9(7), e102108.

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Bacterial Growth Media Preparation

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Luria-Bertani (LB) broth, LB agar and tryptone were obtained from Lab M Limited (Lancashire, United Kingdom). Mueller Hinton broth, Mueller Hinton agar and tryptone soya broth and tryptone soya agar were the products of Oxoid (Hampshire, UK). Metformin hydrochloride, dimethyl sulphoxide (DMSO), N-hexanoyl homoserine lactone and elastin congo red were purchased from Sigma (St. Louis, USA). Other chemicals were of pharmaceutical grade.

Abbas H.A., Elsherbini A.M, & Shaldam M.A. (2017). Repurposing metformin as a quorum sensing inhibitor in Pseudomonas aeruginosa. African Health Sciences, 17(3), 808-819.

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