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Bre­men re­search­ers cul­tiv­ate ar­chaea that break down crude oil in novel ways

Jun 1, 2023
How mi­croor­gan­isms deep in the seabed render crude oil harm­less

The seafloor is home to around one-third of all the microorganisms on the Earth and is inhabited even at a depth of several kilometers. Only when it becomes too hot does the abundance of microorganisms appear to decline. But how, and from what, do microorganisms in the deep seafloor live? How do their metabolic cycles work and how do the individual members of these buried communities interact? Researchers at MARUM – Center for Marine Environmental Sciences at the University of Bremen and at the Max Planck Institute for Marine Microbiology at Bremen have now been able to demonstrate in laboratory cultures how small, liquid components of crude oil are broken down through a new mechanism by a group of microorganisms called archaea. Their results have now been published in the professional journal Nature Mi­cro­bi­o­logy.

Mi­cro­bial com­munit­ies are es­pe­cially act­ive near hy­dro­thermal seeps like those in the Guay­mas Basin in the Gulf of Cali­for­nia. The team of re­search­ers has been work­ing on un­der­stand­ing these com­munit­ies for many years. Or­ganic ma­ter­ial de­pos­ited in the Guay­mas Basin is cooked by heat sources from within the Earth, which breaks it down into crude oil and nat­ural gas. Their com­pon­ents provide the primary source of en­ergy for mi­croor­gan­isms in an oth­er­wise hos­tile en­vir­on­ment. In their latest study, the re­search­ers have demon­strated that ar­chaea use a pre­vi­ously un­known mech­an­ism to de­grade li­quid pet­ro­leum al­kanes at high tem­per­at­ures without the pres­ence of oxy­gen.

Al­kanes are highly stable com­pounds of car­bon and hy­dro­gen. They are nat­ural com­pon­ents of nat­ural gas and crude oil. The lat­ter is re­fined by hu­mans into fuels like gas­ol­ine and ker­osene. En­vir­on­mental cata­strophes oc­cur re­peatedly due to ac­ci­dents dur­ing the ex­trac­tion of crude oil. A prime ex­ample was the ac­ci­dent on the Deep­wa­ter Ho­ri­zon drilling plat­form, which caused severe en­vir­on­mental dam­age in the Gulf of Mex­ico due to the toxic ef­fects of crude oil com­pounds like li­quid al­kanes. In the pres­ence of oxy­gen, mi­croor­gan­isms can rap­idly break down many com­pon­ents of crude oil, among oth­ers al­kanes. Without the re­act­ive oxy­gen, however, de­grad­a­tion is con­sid­er­ably more dif­fi­cult. Or­gan­isms that can per­form this task have not been ex­tens­ively re­searched. In re­cent years, however, evid­ence has been found that ar­chaea are able to em­ploy a sur­pris­ing mech­an­ism to do this. It is based on newly dis­covered vari­ants of the key en­zyme of meth­ano­gen­esis and an­aer­obic meth­ane de­grad­a­tion, methyl-coen­zyme M re­ductase (MCR). The genes that en­code these en­zymes have been found in many en­vir­on­mental samples. However, labor­at­ory cul­tures of the mi­crobes that could il­lus­trate the func­tion of these en­zymes were still lack­ing. This is where the labor­at­ory study of Hanna Zehnle and her col­leagues be­comes sig­ni­fic­ant.

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In the spotlight of the U.S. deep-sea submersible ALVIN, a small reddish-brown vent massif can be seen on the seafloor of the Guaymas Basin. This formation is surrounded by abundant hydrothermally heated oil-rich sediments covered by white and orange bacterial mats. The core from which the Candidatus Alkanophaga archaea ultimately originated was collected by the team of the manned deep-sea submersible. Photo: Woods Hole Oceanographic Institution. © Woods Hole Oceanographic Institution

The team used sed­i­ments from the 2000-meter-deep Guay­mas Basin in the Gulf of Cali­for­nia. The spe­cial geo­lo­gical con­di­tions that ex­ist here in­clude high tem­per­at­ures, li­quid crude-oil com­pon­ents, and an an­aer­obic en­vir­on­ment at shal­low sed­i­ment depths, all of which are nor­mally only found in deep-ly­ing oil reser­voirs which are dif­fi­cult for sci­ent­ists to ac­cess.

In the Bre­men labor­at­or­ies the re­search­ers pre­pared cul­tures with li­quid al­kanes and al­lowed them to grow an­aer­obic­ally, i.e., without oxy­gen, at high tem­per­at­ures (70 de­grees Celsius). “After a time,” ex­plains first au­thor Hanna Zehnle, “sulf­ide forms in the cul­tures. This provides evid­ence that they are act­ive.” The com­pos­i­tion of the cul­tures is stud­ied us­ing DNA and RNA samples. “With this method we can find out what or­gan­isms are liv­ing in this sys­tem and which meta­bolic path­ways they are us­ing,” says Zehnle. These in­clude the chem­ical re­ac­tions in which sub­stances are meta­bol­ized. They found ar­chaea of the genus Candidatus Alkanophaga in the cul­tures. These ar­chaea use vari­ants of the MCR for break­ing down the al­kanes. The re­search­ers veri­fied this by tran­scrip­tome data, meas­ure­ment of the en­zyme products, and by demon­strat­ing in­activ­ity of the cul­tures when the en­zyme was in­hib­ited. But the or­gan­isms are not able to de­grade the crude oil alone. Res­pir­a­tion, in the form of sulfate re­duc­tion in this case (be­cause no oxy­gen is present), is car­ried out by bac­teria of the genus Thermodesulfobacterium, which form dense con­sor­tia with the ar­chaea.

Meth­ano­gen­esis is one of the old­est known meta­bolic pro­cesses and is a part of the global car­bon cycle. The labor­at­ory study by Hanna Zehnle and her col­leagues shows that the en­zymes in­volved in this pro­cess can also util­ize li­quid (and thus toxic) hy­dro­car­bons which high­lights the rel­ev­ance of this path­way for the global car­bon cycle.

“Thanks to their newly dis­covered cap­ab­il­it­ies, Al­kan­o­phaga and their re­l­at­ives are tar­get­ing hy­dro­car­bons in oil reser­voirs. The re­main­ing oil be­comes more and more solid and there­fore tends to re­main in the sea­floor,” ex­plains cor­res­pond­ing au­thor Gunter We­gener. “We still have not been able to in­vest­ig­ate any deep oil reser­voirs, but the ar­chaea are cer­tainly an­noy­ing the oil in­dustry with their activ­ity. But they also make an im­port­ant con­tri­bu­tion to the fact that nat­ural oil seeps are rare.”

This study is a part of the re­search within the Cluster of Ex­cel­lence “The Ocean Floor – Earth’s Un­charted In­ter­face”, which is housed at MARUM. Among other re­search areas, in­vest­ig­a­tions are be­ing con­duc­ted here to de­term­ine which mi­croor­gan­isms and en­vir­on­mental con­di­tions provide the ocean floor with the qual­it­ies re­quired to func­tion as a re­actor that has been bal­an­cing the Earth's car­bon cycle since time im­me­morial. Ques­tions re­lat­ing to what the or­gan­isms in the deep bio­sphere in higher tem­per­at­ure ranges live from is one of the core themes of the Cluster.

Kultur
The microbial community was cultivated in a variety of liquid alkanes, here a hexane culture. An oil layer can be observed on the surface. © Hanna Zehnle

Ori­ginal pub­lic­a­tion

Links

Par­ti­cip­at­ing in­sti­tu­tions

  • MARUM – Center for Marine Environmental Sciences at the University of Bremen, Bremen, Germany
  • Max Planck Institute for Marine Microbiology, Bremen, Germany
  • Faculty of Geosciences, University of Bremen, Bremen, Germany
  • Department of Systems Biology, National Centre for Biotechnology – CSIC, Madrid, Spain
  • Max Planck Institute for Multidisciplinary Sciences, Göttingen, Germany
  • Department of Earth, Marine and Environmental Sciences, University of North Carolina at Chapel Hill, NC, USA

MARUM pro­duces fun­da­mental sci­entific know­ledge about the role of the ocean and the sea­floor in the total Earth sys­tem. The dy­nam­ics of the oceans and the seabed sig­ni­fic­antly im­pact the en­tire Earth sys­tem through the in­ter­ac­tion of geo­lo­gical, phys­ical, bio­lo­gical and chem­ical pro­cesses. These in­flu­ence both the cli­mate and the global car­bon cycle, res­ult­ing in the cre­ation of unique bio­lo­gical sys­tems. MARUM is com­mit­ted to fun­da­mental and un­biased re­search in the in­terests of so­ci­ety, the mar­ine en­vir­on­ment, and in ac­cord­ance with the sus­tain­ab­il­ity goals of the United Na­tions. It pub­lishes its qual­ity-as­sured sci­entific data to make it pub­licly avail­able. MARUM in­forms the pub­lic about new dis­cov­er­ies in the mar­ine en­vir­on­ment and provides prac­tical know­ledge through its dia­logue with so­ci­ety. MARUM co­oper­a­tion with com­pan­ies and in­dus­trial part­ners is car­ried out in ac­cord­ance with its goal of pro­tect­ing the mar­ine en­vir­on­ment.

Please dir­ect your quer­ies to:

Scientist

HGF MPG Joint Research Group for Deep-Sea Ecology and Technology

Dr. Gunter Wegener

MPI for Marine Microbiology
Celsiusstr. 1
D-28359 Bremen
Germany

Room: 

1335

Phone: 

+49 421 2028-8670

Dr. Gunter Wegener

Head of Press & Communications

Dr. Fanni Aspetsberger

MPI for Marine Microbiology
Celsiusstr. 1
D-28359 Bremen
Germany

Room: 

1345

Phone: 

+49 421 2028-9470

Dr. Fanni Aspetsberger
 
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