Two types of extracellular proteases with molecular mass of 50.0 and 44.8 kDa were found in H10 enzymes partially purified from Bacillus cereus H10. Further identification using liquid chromatography-tandem mass spectrometry, the enzyme with 50.0 kDa was identified as being similar to leucine dehydrogenase; while the enzyme with 44.8 kDa might be a novel keratinolytic enzyme with little similarity to other proteins. To maximize the keratinolytic and proteolytic abilities in the H10 enzymes, a combination of response surface methodology and sequential quadratic programming technique was used to study the hydrolytic pH and temperature. Results showed that the H10 enzymes could produce optimal proteolytic and keratinolytic activities at a hydrolysis temperature of 59°C at pH 7.57. Testing the protease activity on various protein substrates and temperatures indicated that the H10 enzymes showed high thermal stability and were very effective in porcine hair.

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hypertrophy), or with elements of CKD-MBD (Vitamin D, iPTH, Ca, P, CaXP) we found no correlation. sKlotho showed a positive correlation with hemodialysis efficiency (eKTV) (r¼0.26, p-0.04), but no correlation with hemodialysis duration. When we classified patients according to median sKlotho level, we found out that patients with sKlotho> 333.398 pg/ml had significantly higher level of hemoglobin (11.2 vs. 10.6 g/dl, p¼0.017). Patients with higher levels of sKlotho showed a decreased risk of mortality, however not statistically significant. We found a higher risk of all cause mortality in patients with diabetes mellitus (OR 6.9, 95% CI-1.6924 to 28.1469, p¼0.007) and with left ventricular hypertrophy (OR 12.4, 95% CI-1.5135 to 102.9073, p¼0.01). Conclusions: In our study, higher levels of sKlotho were associated to a better controlled anemia, a better hemodialysis efficiency, the absence of signs of coronary heart disease on ECG, but not with an increased 2 year survival in hemodialysis patients.

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