Keratinolytic microorganisms have become the subject of scientific interest due to their ability to biosynthesize specific keratinases and their prospective application in keratinic waste management. Among several bacterial classes, actinobacteria remain one of the most important sources of keratin-degrading strains, however members of the Micrococcaceae family are rarely scrutinized in regard to their applicatory keratinolytic potential. The tested Micrococcus sp. B1pz isolate from poultry feather waste was identified as M. luteus. The strain, grown in the medium with 1–2% chicken feathers and a yeast extract supplement, produced keratinases of 32 KU and lower level of proteases, 6 PU. It was capable to effectively decompose feathers or “soft” keratin of stratum corneum, in contrast to other “hard” hair-type keratins. The produced keratinolytic enzymes were mainly a combination of alkaline serine or thiol proteases, active at the optimum pH 9.4, 55 °C. Four main protease fractions of 62, 185, 139 and 229 kDa were identified in the crude culture fluid. The research on the auxiliary role of reducing factors revealed that reducing sulfur compounds could be applied in keratinolysis enhancement during enzymatic digestion of keratin, rather than in culture conditions. The presented M. luteus isolate exhibits a significant keratinolytic potential, which determines its feasible applicatory capacity towards biodegradation of poultry by-products or formulation of keratin-based feed components.

Background:Extensive quantities of keratinic by-products are disposed annually by animal-processing industry, causing a mounting ecological problem due to extreme resilience of these materials to enzymatic breakdown. There is a growing trend to apply cheap and environment-friendly methods to recycle keratinic wastes. Soil bacteria of profound keratinolytic potential, especially spore-forming rods from the genus Bacillus, play a significant role in keratinase-mediated biodegradation of keratins, therefore could be effective in hastening their biodegradation. Keratin hydrolysis in microbial cultures is one of the most promising techniques not only to utilize this protein but also to obtain valuable by products.Objectives:The study was undertaken to investigate the biodegradation process of various keratinic materials by two Bacillus strains.Materials and Methods:Two keratinolytic strains, Bacillus cereus and B. polymyxa, were subject to cultures in the presence of several keratinic appendages, like chicken feathers, barbs and rachea of ostrich feathers, pig bristle, lamb wool, human hair and stratum corneum of epidermis, as main nutrient sources. Bacterial ability to decompose these waste materials was evaluated, at the background of keratinase and protease biosynthesis, in brief four-day cultures. Keratinolytic activity was measured on soluble keratin preparation and proteases were assayed on casein. Additionally, amounts of liberated proteins, amino acids and thiols were evaluated. Residual keratin weight was tested afterwards.Results:Both tested strains proved to be more adapted for fast biodegradation of feather β-keratins than hair-type α-keratins. B. cereus revealed its significant proteolytic potential, especially on whole chicken feathers (230 PU) and stratum corneum (180 PU), but also on separated barbs and rachea, which appeared to be moderate protease inducers. Keratinolytic activity of B. cereus was comparable on most substrates and maximum level obtained was 11 KU. B. polymyxa was found to be a better producer of keratinases, up to 32 KU on chicken feathers and 14 KU on both fractions of ostrich feathers. Its proteolytic activity was mostly revealed on stratum corneum and human hair. Stratum corneum was extensively degraded by both bacterial strains up to 99% - 87%, chicken feathers 47-56%, ostrich barbs and rachea, 28% and 35% at maximum, respectively. Keratin fibres of structures like human hair, lamb wool and pig bristle remained highly resilient to this short microbiological treatment, however certain extent of keratinase induction was also observed.Conclusions:The obtained results prove that keratinolytic potential of both tested bacterial strains could be applied mainly in biodegradation of feathers, however, B. cereus and B. polymyxa differed in terms of keratinase and protease production on each of the substrates. Biodegradation of highly resilient structures like hair or pig bristle requires further analysis of process conditions.

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The aim of this study was to determine the impact of ABCB1 polymorphism, BMI, age and drug co-administration on safety and efficiency of posaconazole (PCZ) oral suspension treatment in children with hematological diseases. Seventy children were included in the study. ABCB1 polymorphism in fifty-eight children was determined using a PCR–RFLP method. A protocol with data on the health condition, treatment and adverse events (AE), as well as a survey on treatment tolerance for the legal guardians was evaluated. Liver function tests were observed for the first 20 days, and AE during the complete medication period. For statistical analysis a χ2 test with Yates’s correction, a Pearson’s or Spearman’s correlation test was performed (p < 0.05). Genetic testing showed 24% CC, 46% CT and 30% of TT variants. PCZ prophylaxis failed in twenty cases, where change in prophylactic treatment was needed. Fifty-two children suffered from at least one mild to moderate adverse event. Sixty-five legal guardians completed the survey, most of them reported the treatment to be well tolerated. ABCB1 polymorphism had no impact on AE occurrence and posaconazole prophylaxis efficiency. Age influenced the number of gastrointestinal (p = 0.02), visual (p = 0.05), neurological (p = 0.01), dermatological (p = 0.002) and flu-like (p = 0.02) complications. AST (p = 0.03) and LDH (p = 0.008) activity presented age dependency. The concomitant use of proton pump inhibitors (PPI) had impact on liver health parameters elevation (p = 0.009) and circulatory system complications (p = 0.008). High incidence of mild to moderate AE, and other factors influencing PCZ pharmacokinetics (PPI co-administration, obesity), suggest a need for careful pediatric onco-hematology patient evaluation.

W hodowli bakterii Bacillus cereus B5e/sz na podłożu syntetycznym z dodatkiem piór kurzych biodegradacji ulegały keratyny zaliczane do białek włókienkowych. Ich hydroliza zachodziła przy udziale wydzielanych do środowiska keratynolitycznych proteaz. Enzymy te upłynniały natywną keratynę w postaci nierozpuszczalnej, a także modyfikowaną chemicznie oraz w znacznie większym stopniu keratynę rozpuszczalną. Bakteryjne keratynazy stanowiły mieszaninę względnie termostabilnych, obojętnych metaloproteaz o aktywności keratynolitycznej 10,9 JK i proteolitycznej 90,5 JP. Metodą in silico wykazano, że w sekwencji aminokwasowej keratyny piór kurzych zawarte są krótkie, bioaktywne peptydy z przewagą inhibitora ACE. Hydrolizat keratyny może być źródłem uzyskiwania peptydów o potencjalnym wykorzystaniu w żywności. Słowa kluczowe: keratyna, keratynoliza, Bacillus cereus, proteazy, bioaktywne peptydy Wprowadzenie Keratyna jest białkowym komponentem pierza stanowiącym 90 % ich masy, a także składnikiem budulcowym innych wyrostków skóry kręgowców: włosów, sierści, pazurów itp. Jest to białko włókienkowe odporne na czynniki fizyczne i chemiczne oraz na działanie enzymów proteolitycznych pochodzenia zwierzęcego [12, 14]. Jego proteoliza możliwa jest z udziałem drobnoustrojowych proteaz specyficznych względem keratyn. Wśród mikroorganizmów mających uzdolnienia keratynolityczne dominującą grupę stanowią bakterie z rodzaju Bacillus: B. subtilis, B. licheniformis, B. pumilis, B. cereus i inne [6].

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