Biotransformation of sulfonamide antibiotics

by ammonia oxidizers

Presenter: Yaochun Yu

1

5/21/2019

19

University of Illinois at Urbana-Champaign

2

Introduction

Objectives

Results

Summary

Outline

3

SAs as one of the biggest families of antibiotics are widely used as human and

animal drugs.

Introduction Objectives Results Summary

Sulfonamide (SA) antibiotics

Why do we care about sulfonamides?

Adverse effects on ecosystem and public health.

- Induce the antibiotic resistance

Sulfonamides functional group

4

Introduction Objectives Results Summary

How do SAs enter into the ecosystem?

https://environmicrobe.weebly.com/research.html

SAs &Unknown transformation products

WWTPs

5

Introduction Objectives Results Summary

Biotransformation of SAs by activated sludge.

6

Activated sludge

Heterotrophs

Introduction Objectives Results Summary

Biotransformation of SAs by activated sludge.

Ammonia oxidizers

Nitrifiers

Nitrite oxidizers

7

Activated sludge

Heterotrophs

Introduction Objectives Results Summary

Biotransformation of SAs by activated sludge.

Ammonia oxidizersAOB

Nitrifiers

Nitrite oxidizer

Yu Y, et al. 2018

Mia

nser

in

Indo

met

hacin

Ran

itidi

ne

Asula

m

Furos

emid

e

Fenhe

xam

id

Ruf

inam

ide

Bezaf

ibra

te

Isop

rotu

ron

Car

bend

azim

Trimet

hopr

im

Mon

uron

Clo

maz

one

Aceta

mip

rid

Leve

tirac

etam

Irgar

ol

Tebuf

enoz

ide

-40

-20

0

20

40

60

80

100 *

16

8h

re

moval (%

)

AOB/NOB co-culture

NOB pure culture

Fresh medium ctrl

Heat-inactivaed ctrl

* *

**

* *

*

8

Activated sludge

Heterotrophs

Introduction Objectives Results Summary

Biotransformation of SAs by activated sludge.

Ammonia oxidizers

AOB AOA Comammox

Nitrifiers

Nitrite oxidizer

Ammonia

Oxidizing

Bacteria

Ammonia

Oxidizing

Archaea

Complete

Ammonia

Oxidizers

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Introduction Objectives Results Summary

Investigate the roles of 3 ammonia oxidizers in SAs biotransformation.

Q1: Can ammonia oxidizers biotransform any SAs?

Q2: What are the transformation mechanisms?

Q3: What are the transformation products?

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AOB: Nitrosomonas nitrosa Nm90

AOA: Nitrososphara gargensis

COMAMMOX: Nitrospira inopinataBatch reactor, triplicates

Introduction Objectives Results Summary

Seven sulfonamides:

Sulfadiazine, sulfadoxine, sulfamethazine, sulfamethoxazole,

sulfamonomethoxine, sulfamerazine, sulfathiazole.

Q1: Can ammonia oxidizers biotransform selected SAs?

AOAAOB

COMAMMOX

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Q1: Can ammonia oxidizers biotransform selected SAs?

A1: Yes, sulfonamides can be biotransformed by ammonia oxidizers.

Introduction Objectives Results Summary

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AOA:N. gargensis

0 100 200 300 400 500

0.2

0.4

0.6

0.8

1.0

0

C/C

0

Time (h)

w/ 0.2 uM ammonia

w/ unlimited ammonia

Sulfadiazine

0.2

0.4

0.6

0.8

1.0

0

C/C

0

Sulfamethazine0 100 200 300 400 500

0.2

0.4

0.6

0.8

1.0

0

C/C

0

Time (h)

0.2

0.4

0.6

0.8

1.0

0

C/C

0

Sulfadiazine

AOB: N. nitrosa Nm90

0.2

0.4

0.6

0.8

1.0

0

C/C

0

Sulfamethazine0 20 40 60 80 100 120 140 160 180

0.2

0.4

0.6

0.8

1.0

0

C/C

0

Time (h)0 50 100 150 200 250 300

0.2

0.4

0.6

0.8

1.0

0

C/C

0

Time (h)

0.2

0.4

0.6

0.8

1.0

0

C/C

0

Sulfamethazine

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

0

C/C

0

Sulfadiazine

COMAMMOX:N. inopinata

Q2: What are the transformation mechanisms?

Introduction Objectives Results Summary

A2: Cometabolism with ammonia oxidation.

(Take sulfadiazine & sulfamethazine as examples)

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Introduction Objectives Results Summary

Q3: What are the transformation products?

A3: Suspect and non-target screening.

TPs of SAs formed activated sludge

Achermann S, et al. 2018

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Introduction Objectives Results Summary

Q3: What are the transformation products?

A3: Suspect and non-target screening.

Deamination (-NH)Sulfadiazine TP236

Pterin conjugation

Pe

ak a

rea

Pe

ak a

rea

Pea

k a

rea

AOA AOB COMAMMOX

For AOB

Pterin-SDZ

TP280

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Introduction Objectives Results Summary

Q3: What are the transformation products?

A3: Suspect and non-target screening.

Pe

ak a

rea

Pe

ak a

rea

Pe

ak a

rea

AOA AOB COMAMMOX

Sulfamethazine

TP264

TP325Pterin-SMZ (AOB)

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Introduction Objectives Results Summary

Ammonia oxidizers

AOB AOA Comammox

Yu Y, et al. 2018

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Introduction Objectives Results Summary

Sulfa

diaz

ine

Sulfa

doxine

Sulfa

met

hazine

Sulfa

met

hoxa

zole

Sulfa

mon

omet

hoxine

Sulfa

mer

azine

Sulfa

thiazo

le0

20

40

60

80

1001

68

h R

em

ova

l (%

)

Abiotic: sulfonamides + hydroxylamine (5μM/12h)

All selected SAs can react with hydroxylamine.

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Introduction Objectives Results Summary

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Introduction Objectives Results Summary

https://doi.org/10.1016/j.watres.2019.05.031

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We thank Dr. Zhong (Lucas) Li form Roy J.

Carver Biotechnology Center.

Dr. Yujie Men

Dr. Michael Wagner

Dr. Ping Han

Dr. Li-Jun Zhou

Acknowledgements

Dr. Baozhan Wang

Dr. Qinglong Wu

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Thanks!

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SAs as one of the biggest families of antibiotics are widely used as human and

animal drugs.

Introduction Objectives Results Summary

Sulfonamide (SA) antibiotics

Sulfonamides functional group

Dihydropteroate

Synthase

(enzyme DHPS)

Folate/

Vitamin B9

Cell growth

How does SA work?