Extractive broth

A 500 mL flask containing 100 mL of the malt extract medium was inoculated with 2 mL frozen stock culture of Streptomyces sp. SC 15761 and incubated for 3 days at 28 °C and 250 rpm. Then 1 mL of the resulting culture was added to each 500 mL flask (11 in total) containing 100 mL of the malt extract broth. The cultures were incubated at 28 °C and 250 rpm for 2 days. Dasatinib (200/rL of a 48.9 mg mL-1 solution in DMSO) was then added to each of the 11 flasks. The flasks were returned to the shaker and incubated for an additional 27 h at 28 °C and 250 rpm. The reaction cultures were pooled and extracted twice, once with 1000 mL of ethyl acetate and once... 

For bacteriological analyses, the commercially available culture media were used Endo agar, elective saline agar, blood agar, thioglycolic medium and beef extract broth (BEB) with 1% glucose. Chocolate agar was used for the isolation of haemophilic flora. 

PDY (Potato Dextrose Yeast) Agar the filtered, extracted broth from boiling 300 grams of sliced potatoes in 1 liter of water for 1 hour20gramsagar... 

Meaty or broth-like (aka yeasty) Yeast extract, meat extract, broth, old yeast. -... 

Autoclavable sugars Glucose (dextrose) and galactose are prepared as a 2% (wt/vol) solution in yeast extract broth described above. When fully solubilized, transfer 10 mL to capped test tubes. 

Transfer 4-mL aliquots of yeast extract broth to test tubes, insert Durham vial (open-side down) into tubes, cap (see Step 2) and autoclave. 

Once yeast extract broth has cooled to room temperature, asepti-cally transfer 2-mL aliquots of each sterile-filtered sugar to broth. 

Among the physiological tests utilized in identification is the ability to ferment carbohydrates. This property can be visualized by the formation and, subsequently, collection of gas in inverted Durham tubes (see Fig. B-1). The medium uses 0.5% (wt/vol) yeast extract broth as a basal medium into which the carbon source of interest is incorporated. The inverted Durham tube is then placed into the medium, the tube capped, and then autoclaved. It is not necessary to attempt to remove the air bubble (s) in Durham vials. During the autoclave cycle, the gas will be displaced. 

Acetic acid is produced from complex basal medium (eg., peptone-yeast extract broth). 

COMPLEX BASAL MEDIUM CATABOLISM (E.G., PEPTONE YEAST EXTRACT BROTH)... 

Prepare yeast extract broth by dissolving 0.5% w/v yeast extract in distilled water. 

For sugars that can be autoclaved (e.g., glucose and galactose), prepare as 2% w/v solutions in the yeast extract broth and transfer 10mL to capped 18 X 150 mm test tubes. Insert a Durham tube open-side down into each tube, loosely replace the cap, and autoclave at 121 C/250 F for 15 min. 

Because many sugars should not be autoclaved, prepare as 2% w/v solutions in the yeast extract broth and individually filter sterilize the solutions through 0.45 im membranes. For raffinose, prepare a 4% w/v solution because some strains only use part of the molecule (Yarrow, 1998). Transfer 10 mL aliquots of yeast extract broth to sterile 18 x... 

An increase in production of laccase and MnP by T. trogii was observed by the addition of easily available carbon and nitrogen sources to the culture medium, such as malt-extract and peptone [90], This is due to the presence of aromatic amino acids tryptophan and tyrosine in the Malt-extract broth. Same observation in Phlebia radiate and Phlebia fascicularia with laccase and MnP activity in T. versicolor of about 2.40 and 2.00 U/mL, respectively was observed [5]. A large increase in LiP production when adding tryptophan to the cultures of T. versicolor, P. chrysosporium and Chrysosporium lignorum was observed [23]. 

Antibiotics. Solvent extraction is an important step in the recovery of many antibiotics (qv) such as penicillin [1406-05-9] streptomycin [57-92-17, novobiocin [303-81-1J, bacitracin [1405-87-4] erythromycin, and the cephalosporins. A good example is in the manufacture of penicillin (242) by a batchwise fermentation. Amyl acetate [628-63-7] or -butyl acetate [123-86-4] is used as the extraction solvent for the filtered fermentation broth. The penicillin is first extracted into the solvent from the broth at pH 2.0 to 2.5 and the extract treated with a buffet solution (pH 6) to obtain a penicillin-rich solution. Then the pH is again lowered and the penicillin is re-extracted into the solvent to yield a pure concentrated solution. Because penicillin degrades rapidly at low pH, it is necessary to perform the initial extraction as rapidly as possible for this reason centrifugal extractors are generally used. 

Biopolymer Extraction. Research interests involving new techniques for separation of biochemicals from fermentation broth and cell culture media have increased as biotechnology has grown. Most separation methods are limited to small-scale appHcations but recendy solvent extraction has been studied as a potential technique for continuous and large-scale production and the use of two-phase aqueous systems has received increasing attention (259). A range of enzymes have favorable partition properties in a system based on a PGE—dextran—salt solution (97) ... 

The batch and fed-batch procedures are used for most commercial antibiotic fermentations. A typical batch fermentor may hold over 150,000 Hters. When a maximum yield of antibiotic is obtained, the fermentation broth is processed by purification procedures tailored for the specific antibiotic being produced. Nonpolar antibiotics are usually purified by solvent extraction procedures water-soluble compounds are commonly purified by ion-exchange methods. Chromatography procedures can readily provide high quaHty material, but for economic reasons chromatography steps are avoided if possible. 

Recovery and Purification. The dalbaheptides are present in both the fermentation broth and the mycelial mass, from which they can be extracted with acetone or methanol, or by raising the pH of the harvested material, eg, to a pH of 10.5—11 for A47934 (16) (44) and A41030 (41) and actaplanin (Table 2) (28). A detailed review on the isolation of dalbaheptides has been written (14). Recovery from aqueous solution is made by ion pair (avoparcin) or butanol (teicoplanin) extraction. The described isolation schemes use ion-exchange matrices such as Dowex and Amberlite IR, acidic alumina, cross-linked polymeric adsorbents such as Diaion HP and Amberlite XAD, cation-exchange dextran gel (Sephadex), and polyamides in various sequences. Reverse-phase hplc, ion-exchange, or affinity resins may be used for further purification (14,89). 

Isolation. Isolation procedures rely primarily on solubiHty, adsorption, and ionic characteristics of the P-lactam antibiotic to separate it from the large number of other components present in the fermentation mixture. The penicillins ate monobasic catboxyHc acids which lend themselves to solvent extraction techniques (154). Pencillin V, because of its improved acid stabiHty over other penicillins, can be precipitated dkecdy from broth filtrates by addition of dilute sulfuric acid (154,156). The separation process for cephalosporin C is more complex because the amphoteric nature of cephalosporin C precludes dkect extraction into organic solvents. This antibiotic is isolated through the use of a combination of ion-exchange and precipitation procedures (157). The use of neutral, macroporous resins such as XAD-2 or XAD-4, allows for a more rapid elimination of impurities in the initial steps of the isolation (158). The isolation procedure for cephamycin C also involves a series of ion exchange treatments (103). 

Pharmaceuticals. Pharmaceuticals account for 6% of the Hquid-phase activated carbon consumption (74). Many antibiotics, vitarnins, and steroids are isolated from fermentation broths by adsorption onto carbon foUowed by solvent extraction and distillation (82). Other uses in pharmaceutical production include process water purification and removal of impurities from intravenous solutions prior to packaging (83). 

Fleisch-saft, m. meat juice, extract of meat, -seite, /. flesh side, -tee, m. beef tea. -ver-giftung, /. meat poisoning, -waren, f.pl. meats, esp. dried meats, -wasser, n. meat broth, -zucker, m. inositol, inosite. 

A selected strain o1 Streptomyces halstedii was cultivated in an aqueous nutrient medium under aerobic conditions and the resulting broth containing carbomycin antibiotics was filtered. The solutions was extracted twice at pH 6.5 with one-quarter volume of methyl isobutyl ketone. The combined extracts were concentrated to one-tenth volume under vacuum, and the antibiotics were extracted into water adjusted to a pH of about 2 with sulfuric acid. After adjusting the separated aqueous solution to pH 6.5, the antibiotic was extracted into benzene and the solution was concentrated to a small volume. Addition of hexane resulted in the separation of a solid product containing the benzene complexes of carbomycin A and carbomycin B, present in the fermentation broth. 

After the end of the fermentation (28 hours) the culture broth is filtered off by suction over a large suction filter. The mycel residue is washed with water several times. The filtrate is extracted three times, each time with 10 liters of methyl isobutyl ketone. The extract is concentrated under vacuum in a circulating evaporator and in a round flask carefully dried under vacuum. The residue is crystallized from acetone/isopropyl ether. The melting point is 157°-158°C (fermentation yield = 60%). The pure product yield obtained after a second crystallization and chromatography of the mother liquor on silica gel amounts to 53% of the theoretical. 

The culture broth (about 1,100 gallons in volume) is adjusted to pH 9.5 with 40% sodium hydroxide solution and is filtered to remove the mycelium, the filtration being assisted by use of 3% of Hyflo Super-Cel, a filter aid, (sold by Johns-IVlanville Company). The clear filtrate is extracted with amyl acetate in a Podbielniak extractor using a ratio of 1 volume of amyl acetate to 6 volumes of clarified broth. The amyl acetate extract is in turn extracted batchwise with water brought to about pH 5 by the addition of sulfuric acid. Two... 

To 100 ml of a sterile nutrient broth (composed of Bacto-beef extract, 3 g Bacto-peptone, 5 g per liter of tap water) in a 300 ml flask is added one loopful of the incubated culture and the broth mixture is further incubated for 24 hours at 28°C on a shaking machine. 

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