Production and characterization of a new detergent-stable keratinase expressed by Pedobacter sp. 3.14.7, a novel Antarctic psychrotolerant keratin-degrading bacterium.

BACKGROUND: Antarctica is one of the harshest environments in the world. Despite this fact, it has been colonized by microorganisms, which had to develop different adaptations in order to survive. By studying their enzymes, we can harness these adaptations in order to use them in various industrial processes. Keratinases (E.C. 3.4.99.11) are characterized by their robustness in withstanding extreme conditions and, along with other enzymes, are commonly added to laundry detergents, which makes their study of industrial interest. RESULTS: In this work, a novel keratinase producer, Pedobacter sp. 3.14.7 (MF 347939.1), isolated from Antarctic birds' nests, was identified. This psychrotolerant isolate displays a typical psychrotolerant growth pattern, with an optimal temperature of 20 °C (µmax=0.23 h-1). After 238 h, maximum proteolytic (22.00 ± 1.17 U ml-1) and keratinolytic (33.04 ± 1.09 U ml-1) activities were achieved with a feather sample conversion of approximately 85%. The keratinase present in crude extract was characterized as a metalloprotease with a molecular weight of 25 kDa, stable in a wide range of pH, with an optimum pH of 7.5. Optimum temperature was 55 °C. Wash performance at 20 °C using this crude extract could remove completely blood stain from cotton cloth. CONCLUSION: We report a new keratinolytic bacteria from maritime Antarctica. Among its biochemical characteristics, its stability in the presence of different detergents and bleaching agents and its wash performance showed promising results regarding its potential use as a laundry detergent additive.

Antarctic yeasts: potential use in a biologic treatment of textile azo dyes.

We investigated the dye-removal potential of a collection of 61 cold-adapted yeasts from the King George Island, Antarctica, on agar plates supplemented with 100 mg L-1 of several textile dyes; among which isolates 81% decolorized Reactive Black 5 (RB-5), with 56% decolorizing Reactive Orange 16, but only 26% doing so with Reactive Blue 19 and Acid Blue 74. Furthermore, we evaluated the ligninolytic potential using 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic-acid) diammonium salt-, 3,5-dimethoxy-4-hydroxybenzaldehydazine-, or manganese-supplemented plates but detected no activity, possibly due to a dye-removal mechanism involving reductases. The removal kinetics were studied in liquid medium supplemented with 100 mg L-1 of RB-5 in a selection of 9 yeasts. The highest volumetric-removal rates (η) were found for Candida sake 41E (4.14 mg L-1 h-1), Leucosporidium muscorum F20A (3.90 mg L-1 h-1), and Cystofilobasidium infirmominiatum F13E (3.90 mg L-1 h-1). Different UV-Vis spectra were obtained if the dye removal occurred by biodegradation or biosorption/bioaccumulation. L. muscorum F20A was selected to study the dye-removal mechanism of RB-5 and the effect of different chemical and environmental parameters on the process. Optimum dye-removal conditions were obtained with 10 g L-1 of glucose within an initial medium pH range of 5.0 to 6.0. Up to 700 mg L-1 of dye could be removed in 45 h. High-performance liquid chromatography profiles obtained were consistent with a biodegradation of the dye. Phytotoxicity was estimated by calculating the 50%-inhibition concentration (IC50) with Lactuca sativa L. seeds. These findings propose psychrophilic yeasts as a novel environmentally suitable alternative for the treatment of dye-industry wastewaters.

Yeasts from sub-Antarctic region: biodiversity, enzymatic activities and their potential as oleaginous microorganisms.

Various microbial groups are well known to produce a range of extracellular enzymes and other secondary metabolites. However, the occurrence and importance of investment in such activities have received relatively limited attention in studies of Antarctic soil microbiota. Sixty-one yeasts strains were isolated from King George Island, Antarctica which were characterized physiologically and identified at the molecular level using the D1/D2 region of rDNA. Fifty-eight yeasts (belonging to the genera Cryptococcus, Leucosporidiella, Rhodotorula, Guehomyces, Candida, Metschnikowia and Debaryomyces) were screened for extracellular amylolytic, proteolytic, esterasic, pectinolytic, inulolytic xylanolytic and cellulolytic activities at low and moderate temperatures. Esterase activity was the most common enzymatic activity expressed by the yeast isolates regardless the assay temperature and inulinase was the second most common enzymatic activity. No cellulolytic activity was detected. One yeast identified as Guehomyces pullulans (8E) showed significant activity across six of seven enzymes types tested. Twenty-eight yeast isolates were classified as oleaginous, being the isolate 8E the strain that accumulated the highest levels of saponifiable lipids (42 %).

Biodegradation of a keratin waste and the concomitant production of detergent stable serine proteases from Paecilomyces lilacinus.

Paecilomyces lilacinus (LPS 876) efficiently degraded keratin in chicken feather during submerged cultivation producing extracellular proteases. Characterization of crude protease activity was done including its compatibility in commercial detergents. Optimum pH and temperature were 10.0 and 60 °C, respectively. Protease activity was enhanced by Ca²âº but was strongly inhibited by PMSF and by Hg²âº suggesting the presence of thiol-dependent serine proteases. The crude protease showed extreme stability toward non-ionic (Tween 20, Tween 85, and Triton X-100) and anionic (SDS) surfactants, and relative stability toward oxidizing agent (H2O2 and sodium perborate). In addition, it showed excellent stability and compatibility with various solid and liquid commercial detergents from 30 to 50 °C. The enzyme preparation retained more than 95% of its initial activity with solid detergents (Ariel™ and Drive™) and 97% of its original activity with a liquid detergent (Ace™) after pre-incubation at 40 °C. The protective effect of polyols (propylene glycol, PEG 4000, and glycerol) on the heat inactivation was also examined and the best results were obtained with glycerol from 50 to 60 °C. Considering its promising properties, P. lilacinus enzymatic preparation may be considered as a candidate for use in biotechnological processes (i.e., as detergent additive) and in the processing of keratinous wastes.