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\begin{document}

\title{Anaerobic hydrolysis of complex substrates in full-scale aerobic
granular sludge: enzymatic activity determined in different sludge
fractions~}



\author[1]{Sara Toja Ortega}%
\author[1]{Mario Pronk}%
\author[1]{Merle de Kreuk}%
\affil[1]{TU Delft}%


\vspace{-1em}



  
  \date{May 10, 2024}


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\selectlanguage{english}
\begin{abstract}
Complex substrates, like proteins, carbohydrates, and lipids, are major
components of domestic wastewater, and yet their degradation in
biofilm-based wastewater treatment technologies, such as aerobic
granular sludge (AGS), is not well understood. Hydrolysis is considered
the rate-limiting step in the bioconversion of complex substrates, and
as such, it will impact the utilization of a large wastewater COD
(chemical oxygen demand) fraction by the biofilms or granules. To study
the hydrolysis of complex substrates within these types of biomass, this
paper investigates the anaerobic activity of major hydrolytic enzymes in
the different sludge fractions of a full-scale AGS reactor. Chromogenic
substrates were used under fully mixed anaerobic conditions to determine
lipase, protease, \selectlanguage{greek}α-\selectlanguage{english}glucosidase, and \selectlanguage{greek}β-\selectlanguage{english}glucosidase activities in large
granules (\textgreater{}1 mm in diameter), small granules (0.2--1 mm),
flocculent sludge (0.045--0.2 mm), and bulk liquid. Furthermore,
composition and hydrolytic activity of influent wastewater samples were
determined. Our results showed an overcapacity of the sludge to
hydrolyze wastewater soluble and colloidal polymeric substrates. The
highest specific hydrolytic activity was associated with the flocculent
sludge fraction (1.5--7.5 times that of large and smaller granules), in
agreement with its large available surface area. However, the biomass in
the full-scale reactor consisted of 84\% large granules, making the
large granules account for 55--68\% of the total hydrolytic activity
potential in the reactor. These observations shine a new light on the
contribution of large granules to the conversion of polymeric COD and
suggest that large granules can hydrolyze a significant amount of this
influent fraction. The anaerobic removal of polymeric soluble and
colloidal substrates could clarify the stable granule formation that is
observed in full-scale installations, even when those are fed with
complex wastewaters.%
\end{abstract}%



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