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Enzymes catalyze key reactions within Earth’s life-sustaining biogeochemical cycles. Here, we use metaproteomics to examine the enzymatic capabilities of the microbial community (0.2 to 3 µm) along a 5,000-km-long, 1-km-deep transect in the central Pacific Ocean. Eighty-five percent of total protein abundance was of bacterial origin, with Archaea contributing 1.6%. Over 2,000 functional KEGG Ontology (KO) groups were identified, yet only 25 KO groups contributed over half of the protein abundance, simultaneously indicating abundant key functions and a long tail of diverse functions. Vertical attenuation of individual proteins displayed stratification of nutrient transport, carbon utilization, and environmental stress. The microbial community also varied along horizontal scales, shaped by environmental features specific to the oligotrophic North Pacific Subtropical Gyre, the oxygen-depleted Eastern Tropical North Pacific, and nutrient-rich equatorial upwelling. Some of the most abundant proteins were associated with nitrification and C1 metabolisms, with observed interactions between these pathways. The oxidoreductases nitrite oxidoreductase (NxrAB), nitrite reductase (NirK), ammonia monooxygenase (AmoABC), manganese oxidase (MnxG), formate dehydrogenase (FdoGH and FDH), and carbon monoxide dehydrogenase (CoxLM) displayed distributions indicative of biogeochemical status such as oxidative or nutritional stress, with the potential to be more sensitive than chemical sensors. Enzymes that mediate transformations of atmospheric gases like CO, CO 2 , NO, methanethiol, and methylamines were most abundant in the upwelling region. We identified hot spots of biochemical transformation in the central Pacific Ocean, highlighted previously understudied metabolic pathways in the environment, and provided rich empirical data for biogeochemical models critical for forecasting ecosystem response to climate change. 

Abstract. Large-scale climatic forcing is impactingoceanic biogeochemical cycles and is expected to influence the water-columndistribution of trace gases, including methane and nitrous oxide. Our abilityas a scientific community to evaluate changes in the water-column inventoriesof methane and nitrous oxide depends largely on our capacity to obtain robustand accurate concentration measurements that can be validated acrossdifferent laboratory groups. This study represents the first formalinternational intercomparison of oceanic methane and nitrous oxidemeasurements whereby participating laboratories received batches of seawatersamples from the subtropical Pacific Ocean and the Baltic Sea. Additionally,compressed gas standards from the same calibration scale were distributed tothe majority of participating laboratories to improve the analytical accuracyof the gas measurements. The computations used by each laboratory to derivethe dissolved gas concentrations were also evaluated for inconsistencies(e.g., pressure and temperature corrections, solubility constants). Theresults from the intercomparison and intercalibration provided invaluableinsights into methane and nitrous oxide measurements. It was observed thatanalyses of seawater samples with the lowest concentrations of methane andnitrous oxide had the lowest precisions. In comparison, while the analyticalprecision for samples with the highest concentrations of trace gases wasbetter, the variability between the different laboratories was higher:36% for methane and 27% for nitrous oxide. In addition, thecomparison of different batches of seawater samples with methane and nitrousoxide concentrations that ranged over an order of magnitude revealed theramifications of different calibration procedures for each trace gas.Finally, this study builds upon the intercomparison results to developrecommendations for improving oceanic methane and nitrous oxide measurements,with the aim of precluding future analytical discrepancies betweenlaboratories.