Biosurfactant producing biofilms for the enhancement of nitrification and subsequent aerobic denitrification

Abstract

Wastewater treatment methods have always gravitated towards the use of biological methods for the treatment of domestic grey water. This has been proven to offer a series of advantages such as the reduction of pollution attributed to the use of synthetic chemicals; therefore, this decreases the requirement of further costly post primary treatment methods. Although such biological methods have been used for decades, their efficiency and sustainability has always been challenged by inhibitory toxicants which renders the systems redundant when these toxins are prevalent in high concentrations, culminating in the deactivation of biomass which facilitates the treatment. In most instances, this biomass is anaerobic sludge. Hence, the proposal to utilize biofilms which are ubiquitous and selfsustaining in nature. The use of engineered biofilms in wastewater treatment and their behaviour has been studied extensively, with current research studies focusing on reducing plant footprint, energy intensity and minimal usage of supplementary synthetic chemicals. An example of such processes include traditional nitrification and denitrification systems, which are currently developed as simultaneous nitrification and aerobic denitrification systems, i.e. in a single stage system, from the historical two stage systems. However, there is limited literature on biofilm robustness against a potpourri of toxicants commonly found in wastewater; particularly for total nitrogen removal systems such as simultaneous nitrification and denitrification (SND). This study was undertaken (aim) to assess the ability of biosurfactant producing biofilms in the removal of total nitrogen in the presence of toxicants, i.e. heavy metals and phenol, as biosurfactants have been proven to facilitate better mass transfer for pollutant mitigation. Unlike in conventional studies, the assessment of biosurfactant producers in total nitrogen removal was assessed in both planktonic and biofilm state. Since biofilms are known to have increased tolerance to toxic environmental conditions, they were developed thus engineered using microorganisms isolated from various sources, mainly waste material including wastewater as suggested in literature reviewed, to harness microorganisms’ possessing specified traits that can be developed when organisms are growing under strenuous environments whereby they are tolerant to toxic compounds. The assessment of these engineered biofilms involved the development from individual microorganisms to form biofilms in 1L batch reactors where the isolated microorganisms were grown in basal media containing immobilisation surfaces. The assessment of the total nitrogen efficiency was conducted using Erlenmeyer flasks (500mL) in a shaker incubator, with the biofilm TN removal efficiency being assessed in batch systems to ascertain simultaneous nitrification and denitrification rates even in the presence of heavy metals (Cu2+, Zn2+) and C6H5OH. Ambient temperature and dissolved oxygen conditions were kept constant throughout the duration of biofilm development with microorganisms (initially n = 20) being isolated for the initiation of biosurfactant studies which included screening. Results indicated that the engineered biofilms, constituted by biosurfactant producing organisms (n = 9), were consisiting of bacteria (97.19%), Protozoa (2.81%) and Archaea (0.1%) as identified using metagenomics methods. Some of the biosurfactant produced had the following functional group characteristics as determined by FTIR: -CH3-CH2, deformed NH, -CH3 amide bond, C-O, C=O, O-C-O of carboxylic acids, and C-O-C of polysaccharides. Other selected microorganisms (n = 5) tolerated maximum concentrations of the selected toxicants (Cu2+, Zn2+ and C6H5OH) of 2400 mg/L, 1800 mg/L and 850 mg/L, respectively. Enzyme analysis of the total nitrogen removal experiments indicated a higher nitrogen removal rate to be the Alcanigene sp. at 180 mg/L/h.