Bioreduction of Cr (VI) by potent novel chromate resistant alkaliphilic Bacillus sp. strain KSUCr5 isolated from hypersaline Soda lakes

Abstract

Isolation of Cr (VI) resistant alkaliphilic bacteria from sediment and water samples collected from Wadi Natrun hypersaline Soda lakes (located in northern Egypt), resulted in isolation of several alkaliphilic bacterial strains that can tolerate up to 2.94 g/l of Cr (VI) in alkaline medium. However, with increasing Cr (VI) concentration up to 29.4 g/l, only one strain, KSUCr5, was able to tolerate up to 22 g/l (75 mM) and with MIC value of 23.5 g/l (80 mM) in alkaline medium (pH 10.5) containing 10% NaCl. Based on the 16S rRNA gene analysis, strain KSUCr5 was identified as Bacillus sp. with 99% similarity and was referred to as Bacillus sp. KSUCr5. In addition, Bacillus sp. strain KSUCr5 showed high tolerance to several other heavy metals including Cd²⁺ (50 mM), Mo²⁺ (75 mM), Mn²⁺ (100 mM), Cu²⁺ (2 mM), Ni²⁺ (100 mM), Pb (75 mM), Co²⁺ (5 mM) and Zn²⁺ (2 mM). Strain KSUCr5 was shown to be of a high efficiency in detoxifying chromate, as it could rapidly reduce up to 40 mg/l of Cr(VI) to a non detectable level over 24 h. In addition, at initial Cr(VI) concentration of 60 to 80 and 100 mg/l, 100% of the chromate reduction was achieved within 48 and 72 h, respectively. Strain KSUCr5 could reduce Cr(VI) efficiently  over a wide range of initial Cr(VI) concentrations (10 to 300 mg/l) in alkaline medium under aerobic conditions without significant effect on the bacterial growth. It was able to reduce Cr(VI) in a wide range of NaCl (0 to 20%) with a maximum reduction yield at concentration of 0 to 1.5%, indicating the halo tolerance nature of the bacterium. It was found that addition of glucose and Na₂CO₃ to the culture medium caused a dramatic increase in Cr(VI)-reduction by Bacillus sp. strain KSUCr5. The maximum chromate removal was exhibited in alkaline medium (pH 10) containing 1.2% Na₂CO₃, 1.5% glucose and 1% NaCl and at incubation temperature of 35°C and culture shaking of 150 rpm. Under optimum Cr (VI) reduction conditions, Cr(VI) concentration of 80 mg/l was completely reduced within 24 h, with reduction rate of 3.3 mg h^-1 which is one of the highest Cr(VI) reduction rate under high alkaline conditions, compared with other microorganisms that has been reported so far. Furthermore, the presence of other metals such as Ni²⁺, Mo²⁺, Cu²⁺ and Mn²⁺ at concentration of 100 mg/l together with Cr(VI) in the culture medium slightly increased Cr(VI)-reduction by the strain KSUCr5. Moreover, the isolate, Bacillus sp.  strain KSUCr5, exhibited an ability to repeatedly reduce hexavalent chromium without any amendment of nutrients, suggesting its potential application in continuous bioremediation of Cr(VI). The results reveal the possible isolation of potent heavy metals resistant bacteria from extreme environment such as hypersaline Soda lakes and their application in bioremediation of heavy metals.

Key words: Chromate reduction, bioremediation, heavy metals, Bacillus sp., Soda lakes.