Bacterial lipases

Systemic antibiotics are indicated for moderate-severe inflammatory acne not responding to topical treatments. Systemic antibiotics act on 1) suppression of P. acnes growth 2) inhibition of bacterial lipases 3) reduction of free fatty acids and 4) reduction of inflammation. Oxytetracycline and its derivatives are the most commonly used oral antibiotics. Second-generation tetracyclines such as minocycline, doxy-cycline and lymecycline present longer half-lives, enhanced bacterial activity and lower... 

Jaeger, K.E. and S.Wohlfarth. (1993) Bacterial lipases Biochemistry, molecular genetics and applications in biotechnology. Bioengineering, 9, 39-46. 

Adams, D. M. and Brawley, T. G. 1981. Heat resistant bacterial lipases and ultra-high temperature sterilization of dairy products. J. Dairy Sci. 64, 1951-1957. 

Kouker, G. and Jaeger, K.-E. 1987. Specific and sensitive plate assay for bacterial lipases. Appl. Environ. Microbiol. 53 211-213. 

Stead, 1989). Measurement of lipase activity has received considerable attention because of the defects caused in stored milk and dairy products by low levels of heat-resistant bacterial lipases (Deeth and Touch, 2000 Chen et al., 2003)... 

Lipolysis in milk is affected by inhibiting and activating factors. As discussed above, proteose peptone fraction of milk can inhibit milk LPL while apolipoproteins stimulate the enzyme. This is particularly important in spontaneous lipolysis however, proteose peptone 3 has been shown to inhibit lipolysis induced by homogenization, sonication, and temperature activation (Arora and Joshi, 1994), while protein components of the milk fat globule membrane inhibit lipolysis caused by bacterial lipase (Danthine et al., 2000). Several exogenous chemical agents can also inhibit lipolysis (Collomb and Spahni, 1995). For example, polysaccharides such as X-carrageenan at 0.3 g/1 effectively inhibits lipolysis in milk activated by mechanical means or temperature manipulation (Shipe et al., 1982) and lipolysis caused by the lipase from P. fluorescens (Stern et al., 1988). 

Some progress has been made towards developing a sensitive ELISA method for bacterial lipases (Stepaniak et al., 1987b). However, this technique detects the enzyme protein rather than its activity and hence may be of little practical value to the dairy industry. Further developments can be expected in this field. 

The more common methods used for milk and bacterial lipases are summarized in Table 15.5. 

Tributyrin, olive oil NaCl Continuous potentiometric titration Milk lipase, bacterial lipase Parry et al. (1966), Egelrud and Olivecrona (1973), Andersson et al. (1979), Makhzoum et al. (1996)... 

Tributyrin Triolein NaCl - Bacterial lipase Tomasini et al. (1993)... 

Olive oil/triolein Sonication before and during assay " Bacterial lipase Linfield et al. (1985), Yang et al. (1994)... 

Milk fat Milk fat Blood serum, heparin, CaCL Extraction with isopropanol-heptane-fLSO., titration to visual endpoint Extraction with isopropanol-heptane-fLSO., titration to visual endpoint Milk lipase Bacterial lipase Deeth and Fitz-Gerald (1977) Fitz-Gerald and Deeth (1983)... 

Naphthyl caprylate Thiomersal, bile salts, Ca Extraction of (3-naphthol-Fast Blue complex with ethyl acetate, spectrophotometry at 540 nm Bacterial lipase McKellar and Chollette (1986)... 

ANitrophenyl caprylate Reflectance colorimetry (b color) spectrophotometry at 418 nm Bacterial lipase Blake et al. (1996), Bendicho et al. (2001)... 

ANitrophenyl butyrate (c) Indigo methods Acetonitrile Addition of Clarifying Agent , spectrophotometry at 420 nm Bacterial lipase Humbert et al. (1997)... 

Indoxyl acetate and caprylate 3. Fluorimetric Colorimetry of indigo formed Bacterial lipase Slack (1987), Allen (1994), Munnich and Haasmann (1999), Brand et al. (2000)... 

Umbelliferyl myristate Fluorimetry at 470 nm (emission), 360 nm (excitation) Bacterial lipase de Laborde de Monpezat et al. (1990)... 

Triacylglycerols (esp. triolein) Chloroform-methanol extraction, HPLC with detection at 208 nm GC of derivatized or free fatty acids. Bacterial lipase Veeraragavan (1990), Choi and Jeon (1993)... 

Christen, G.L., Wang, W.-C., Ren, T.-J. 1986. Comparison of the heat resistance of bacterial lipases and proteases and the effect on ultra-high temperature milk quality. J. Dairy Sci. 69, 2769-2778. 

Nadkarni, S.R. 1971. Studies on bacterial lipase. Part II. Study of the characteristics of partially purified lipase from Pseudomonas aeruginosa. Enzymologia 40, 302—313. 

Lipases are the most frequently used enzymes in organic chemistry, catalyzing the hydrolysis of carboxylic acid esters or the reverse reaction in organic solvents [3,5,34,70]. The first example of directed evolution of an enantioselective enzyme according to the principle outlined in Fig. 11.2 concerns the hydrolytic kinetic resolution of the chiral ester 9 catalyzed by the bacterial lipase from Pseudomonas aeruginosa [8], This enzyme is composed of 285 amino acids [32]. It is an active catalyst for the model reaction, but enantioselectivity is poor (ee 5 % in favor of the (S)-acid 10 at about 50 % conversion) (Fig. 11.10) [71]. The selectivity factor E, which reflects the relative rate of the reactions of the (S)- and (R)-substrates, is only 1.1. 

Oral tetracycline or doxycycline can be an effective therapy for noninfections conditions involving the eye such as acne rosacea and meibomianitis. When patients with acne rosacea or meibomianitis receive oral tetracycline, two changes occur amelioration of the symptoms and rednction of free fetty acids in the surfece sebum. Free fetty acids are released from sebum by bacterial lipases and are irritating as well as inflammatory. Tetracycline causes a significant decrease in lipase... 

Suzuki, A., Mizumoto, A., Sarr, M.G. and Dimagno, E. P. (1997) Bacterial lipase and high-fat diets in canine exocrine pancreatic insufficiency a new therapy of steatorrhea Gastroenterology 112, 2048-2055. 

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