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Comparison of methods for detecting protein extracted from excess activated sludge

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Abstract

The protein contents of hydrolyzed sludge supernatant are commonly determined with the Kjeldahl method, but this method suffers from complicated operations, long process times, and large quantities of chemicals consumed. In this paper, the Lowry, bicinchoninic acid (BCA), and Bradford methods were used to test the precision and spiked recovery of proteins from sludge supernatants hydrolyzed by alkaline-thermal hydrolysis (ATH), enzymatic hydrolysis (EH), and ultrasound-assisted enzymatic hydrolysis (UEH), and the results were compared with those obtained with the Kjeldahl method. For all the hydrolytic processes, the sludge protein values determined with the three tested methods were within 0.05 of each other, which met the experimental requirement for accuracy. Both the Lowry and BCA methods had recovery rates of 95–105%, while the Bradford method showed large deviations and was not highly reliable. The three protein determination methods showed significant differences with the Kjeldahl method (P<0.05). However, the relative deviation between the Kjeldahl and BCA methods was the smallest (3–5%), followed by those between the Kjeldahl and the Lowry (11–21%) and Bradford methods (21–90%), and the causes of the deviations were analyzed based on the protein hydrolysate components and the mechanisms for the different detection methods. On the basis of these results, the BCA method was chosen as the most appropriate quantification method for use with sludge protein extraction, and it was used to analyze the protein contents extracted from residual sludge samples obtained from two sewage treatment plants. The reliability of the method was verified, and this lays a foundation for the extraction and reclamation of sludge proteins.

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Funding

This work was supported by the National Key Research and Development Program of China (2021YFD1700900) and the Key Scientific Research Project of Higher Education in Henan Province (Program No. 22A610006).

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Authors and Affiliations

  1. School of Ecology and Environment, Zhengzhou University, Zhengzhou, 450001, China

    Yixin Yan, Mengnan Zhang, Jianlei Gao, Lei Qin, Xi Fu & Junfeng Wan

  2. Henan International Joint Laboratory of Environment and Resources, Zhengzhou University, Zhengzhou, 450001, People’s Republic of China

    Junfeng Wan

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  1. Yixin Yan
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  2. Mengnan Zhang
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  3. Jianlei Gao
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  4. Lei Qin
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  5. Xi Fu
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Contributions

Yixin Yan: conceptualization, formal analysis, writing—review and editing, resources, supervision, funding acquisition, project administration. Mengnan Zhang: investigation, data analysis, visualization, writing—original draft. Jianlei Gao: methodology, supervision, validation. Lei Qin: data curation. Xi Fu: software, writing—original draft. Junfeng Wan: writing—review and editing.

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Correspondence to Jianlei Gao.

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Yan, Y., Zhang, M., Gao, J. et al. Comparison of methods for detecting protein extracted from excess activated sludge. Environ Sci Pollut Res 30, 60967–60975 (2023). https://doi.org/10.1007/s11356-023-26455-x

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