Enzymatic Processes in Marine Biotechnology

In previous review articles the attention of the biocatalytically oriented scientific community towards the marine environment as a source of biocatalysts focused on the habitat-related properties of marine enzymes. Updates have already appeared in the literature, including marine examples of oxidoreductases, hydrolases, transferases, isomerases, ligases, and lyases ready for food and pharmaceutical applications. Here a new approach for searching the literature and presenting a more refined analysis is adopted with respect to previous surveys, centering the attention on the enzymatic process rather than on a single novel activity. Fields of applications are easily individuated: (i) the biorefinery value-chain, where the provision of biomass is one of the most important aspects, with aquaculture as the prominent sector; (ii) the food industry, where the interest in the marine domain is similarly developed to deal with the enzymatic procedures adopted in food manipulation; (iii) the selective and easy extraction/modification of structurally complex marine molecules, where enzymatic treatments are a recognized tool to improve efficiency and selectivity; and (iv) marine biomarkers and derived applications (bioremediation) in pollution monitoring are also included in that these studies could be of high significance for the appreciation of marine bioprocesses.


Introduction
Currently there is enormous interest in marine biotechnology with the worldwide flourishing of editorial initiatives (journals, books, etc.) hosting important experimental results and surveys from several projects, especially those belonging to the FP7 program and to the topic "Blue growth" of H2020, potentially providing clues that will aid development of enabling technologies in the field. The oceans are the world's largest ecosystem, covering more than 70% of the earth's surface. They host the greatest diversity of life in unexplored habitats. The Census of Marine Life, evaluating marine biodiversity, ascertained that at least 50% and potentially more than 90% of marine species are undescribed by science [1]. Only a deep understanding of the complexity of the marine ecosystem will enable human beings to protect the oceans and organisms populating them, and pave the way for sustainable exploitation of marine resources. This knowledge will certainly fuel various applications and itself constitutes the core of marine biotechnology. Increasingly labeled "blue biotechnology", this wide field covers many aspects that are highly relevant to societal challenges, as is well established in the EU Framework Programme Horizon 2020. Several outlined emerging technologies are (i) robotics; (ii) miniaturized solutions for marine monitoring; (iii) biomimetics; (iv) acoustics; (v) nanobiotechnlogy; (vi) renewable energy harvesting (wave energy, algae biofuels); and (vii) high-performance computing. However, many challenges remain, including a deep comprehension of the "marine biotechnology landscape" and a multidisciplinary approach, not only in education and training [2].
Marine sources (microorganisms in general and symbionts in particular, extremophiles, fungi, plants and animals) are of scientific interest in that the origin of marine biomolecules (i.e., biocatalysts) features all marine bioprocesses. Knowledge of these biocatalysts and their habitat-related properties such as salt tolerance, hyperthermostability, barophilicity and cold adaptivity (of great interest for industry) is necessary for bioprocesses exploitation. One of the most explicative aspects is related to the stereo-chemical properties of a marine enzyme. Substrate specificity and affinity, as evolved properties that are linked to the metabolic functions of the enzymes, are key aspects. Two review articles already appeared in 2010 [3] and in 2011 [4] with different focuses on these topics. The first [3], to draw the attention of the biocatalytically oriented scientific community to the marine environment as a source of biocatalysts; in fact the discussion was mainly about the specific diversity of molecular assets of biocatalysis that were recognized with respect to terrestrial counterparts. The second review [4] spotlighted habitat-related properties from a biochemical point of view, also reporting on important examples in bioprocesses. Various updates of these former analyses of the literature have also recently been published, such as the one by Lima et al. 2016 [5] including marine examples of oxidoreductases, hydrolases, transferases, isomerases, ligases and lyases ready for food and pharmaceutical applications.
In the present review, a new approach for searching the literature and presenting a more refined analysis is adopted with respect to previous surveys. The focus of the literature search is centered on the enzymatic process more than on a single novel activity. This survey is developed according to the biotechnological field of applications where bioprocesses, based on marine enzymes and/or marine biomasses, are central. Focusing on enzymatic processes rather than on single activities helsd us to recognize the fields of application. For the first, a biorefinery value-chain, the provision of biomass is one of the most important aspects, with aquaculture as the prominent sector. In the food industry the interest in the marine domain is similarly developed to deal with the enzymatic procedures adopted in food manipulation. Moreover, as for the selective and easy extraction/modification of structurally complex marine molecules, enzymatic treatments are a recognized tool to improve efficiency and selectivity in fine chemistry processes to get access to bioactive compounds and provide complex core blocks ready for hemisynthesis. In closing, the field of marine biomarkers and derived applications (bioremediation) in pollution monitoring could be of high significance for the appreciation of marine bioprocesses.
In the fields indicated above, the selected primary articles are presented in tabulated form, picking up different aspects of importance in short explicative notes to avoid a huge amount of text. Selected modern review articles are listed under each paragraph to depict the present state of the art of the related field.

Literature Search
A survey of the literature has been conducted mainly by using the database ScienceDirect with access to 3800 scientific journals in major scientific disciplines. The search was based on two terms: (i) "enzymatic processing" (in abstract, title or keywords) and (ii) "marine" (in all fields). How to manipulate these queries to get an effective result in terms of the number of hits was first investigated using the search functions offered by the database. In particular, the W/in function (proximity operator) has been found useful for the first query, with the keywords used in the query sorted into phrases (low numbers), sentences (medium) and paragraphs (high) of the hit. This result is then refined by using the AND operator with the word "marine" in all fields. The resulting pattern of these searches is as follows: W/3 (137 hits), W/4 (153), W/5 (161), W/15 (256) and for W/50 (442). As a comparison, a more general coverage alternative search was also used, adopting the same keywords, in the Scopus database. It resulted in a similar score (478 hits), thus confirming the choice of ScienceDirect as the reference database when using W/50. This has been the value adopted for searching for articles for this review.
An interesting detail is the yearly distribution of the hits (shown in Figure 1 below). Two time intervals can be easily recognized; published results are doubled yearly from 2012 to 2016 with respect to the previous score in the interval 1993 to 2011, characterized by fewer than 20 hits per year. This picture reflects the strategic efforts of various funded programs created by the European Commission to support and foster research in the European Research Area and similar actions in other parts of the world. That logical reason for the variation of the results in the two intervals 1993-2011 and 2012-2016 further confirms the suitability of the keywords adopted.

Biorefinery
A future sustainable economy based on renewable resources is the main point of the concept of a biorefinery. Research and development studies have been underway for many years in different parts of the world to replace a large fraction of fossil resources. The most important aspect of a biorefinery value-chain is the provision of biomass with a consistent and regular supply of renewable carbon-based raw materials [6]. One of the sectors providing these feedstocks is aquaculture (algae and seaweeds), which, together with biochemical processes (marine enzymes in pre-treatment) adopted, is an important aspect of the domain of marine biotechnology. Needless to say, the focus in many review articles [7] is on the importance of extremophiles and thermostable enzymes to overcome the limitations of biocatalysts in current bioprocesses for lignocellulosic biomass conversion. In this context, it is also of interest to mention the features of marine biocatalysts related to the ecological features of the habitat in which marine organisms thrive. Generally the resulting enzymatic properties are very important from a biotechnological point of view [4,8].
The idea here is to depict both the state of the art about marine enzyme-based bioprocesses and the importance of marine-originating feedstocks in biorefinery. Therefore, biocatalysts and biomass are the two fundamental elements on which the analysis of primary articles in the literature is based here (Table 1 [ ). The selected articles inserted in Table 1 deal with (i) cellulases and other important carbohydrate-active enzymes; (ii) lipases, to manipulate feedstock oils for biodiesel production and (iii) other biocatalysts, including those commercially available. However, to depict the current state of interest when only marine feedstock exploitation is present, chemical treatments were also listed in these selected modern articles (entries 13-34, Table 1). Non-conventional sources of cellulase enzymes have been sought for a long time. As mentioned in an old report, the identification of these biocatalysts in the marine fungus Dendryphiella arenaria [43] dates back 40 years. However, of recent interest are the complex biopolymers in microalgae cells subjected to breakdown in biological pretreatments, as reported in a more modern review [44]. The focus is not only on cellulases as the most-explored specific and efficient enzymes (entries 1-5 Table 1), but also on other hydrolytic enzymes, including hemicellulase, pectinase, protease and amylase, even in the form of an enzymatic cocktail seen as an effective tool with respect to single enzymes as well. Entire microbial communities associated with marine organisms are studied (entry 2  Table 1). Chitinases are very well represented in research, too, and new insights into the disruption of crystalline polysaccharides were gained, as seen in a recent review report [45]. Substrate-disrupting accessory non-hydrolytic proteins are novel tools to improve molecular accessibility to polymers with increased process efficiency. Other primary articles, dealing with different carbohydrate active hydrolases, are listed in Table 1 (entries [6][7][8].
Obviously lipases are also considered a convenient tool for converting a wide range of feedstock oils into biodiesel [46]. A study of 427 yeast strains from seawater, sediment, mud of salterns, guts of the marine fish and marine algae should also be mentioned here [47]. Industrial yeast Yarrowia lipolytica of marine origin is a biocatalyst of interest in metabolic engineering studies used to expand the substrate range [48]. Entries 9-12 in Table 1 are interesting primary articles along this line of research. Microalgae cultivation and macroalgae developed as pests due to eutrophication are generally seen as potential resources for biofuel production [49]. Another interesting aspect is the combination of macroalgae cultivation exploiting nutrients coming from marine aquaculture or other processes.
In industrial squid manufacturing for chitin production, a large volume of protein effluents containing peptones from alkaline and enzymatic hydrolysis of the pens are substrates used as a nitrogen source to reduce the cost of marine probiotic bacteria cultivation (entry 13, Table 1). A similar approach was also reported for effluents originating from the industrial thermal treatment of mussels (entry 14, Table 1); many other examples on these lines are reported here (entries 15-33, Table 1), where studies on different biomasses were conducted often by comparing chemical and enzymatic procedures and yeast fermentations adopted for bioethanol production. An enzymatic cocktail of glucanase, cellulase and glucosidase was studied for R-phycoerythrin extraction, assisted by ultrasound technology from the red seaweed G. turuturu (entry 34, Table 1) proliferating along the French coast.
Marine biomass-centered studies are also listed in the section about food industry development. Additionally, a particular and interesting aspect studied is the use of algicidal microorganisms to improve cell disruption during biotechnological processes aimed at producing biofuels. Secreted algicidal substances to be used as microalgae breakdown agents are reviewed [50].

Food Applications
Enzymatic procedures in food processing are mostly based on biocatalysts of terrestrial or microbial origin, with new enzymes currently obtained from these environments. For years the marine domain has been seen as a promising source of interesting biocatalysts [51] for modern applications. However, the enzymatic activities present in seafood or in byproducts were utilized for centuries in the traditional production of various cured and fermented seafood, allowing the preparation of numerous sauces and pastes from the time of the Greeks and Romans and also seafood processes in the Far East [52].
The recovery and processing of waste is generally a challenge in the food industry. Especially in the seafood sector, a more complete utilization of the raw material with minimization of the inherent problems of pollution and waste treatment is a current issue. Many reviews or chapters in books depicting the state of the art of this specific topic were found. One of the oldest reports found during our search dates back to 1978; interestingly, it already pointed out bioconversion tools for the processing and valorization of food waste in the conversion to useful products [53]. Upgrading of sea byproducts is, however, still central in more modern analysis [54], with more attention dedicated nowadays to therapeutic potential. Attention to antihypertensive and immunomodulatory agents (i.e., peptides obtained by enzymatic hydrolysis of fish proteins) is recognized more than the simple nutritional and biological properties of these materials. Both have recently been investigated in marine mussels [55]. While for mussel proteins the focus is on peptides obtained by bioprocesses, lipids (PUFAs) are also investigated for the prevention and treatment of rheumatoid arthritis. The use of all-natural stabilizers for food, in the form of (enzymatically) muscle-derived extracts, appears interesting, as well as the addition of plant extracts or pure phenolic compounds to combat oxidation in seafood [56]. Basic research related to the enzymatic processes occurring in seafood material has also been traced; a study of blackening processes in freeze-thawed prawns during storage is of interest. The respiratory pigment hemocyanin is converted into a phenoloxidase-like enzyme and acts as a potent inducer of post-harvest blackening; these discoveries are helpful for the development of anti-blackening treatments for these foods [57]. Biotechnologists are also interested in the large availability of seafood raw materials; byproducts from waste in processing (liver, skin, head, viscera, trimmings, etc.) amount to 60% or more [58] of total renewable raw material. Review articles were found for general aspects [59,60] and a particular focused on seaweeds [61].
The food and biorefinery sections overlap because byproducts, primarily used as feed with low returns, are also thought to be useful for biodiesel generation. Some articles could thus be listed in both sections; however, repetition is avoided here and also in this case tabulation is based on the same fundamental key for the analysis of literature adopted previously in Table 1; enzyme (marine or commercial)-based bioprocesses and types of marine-originating feedstocks are the two columns of Table 2, including a third one for notes . Commercial or marine-originating proteases and lipases are most used for production in this field and a wide range of edible biomass for the valorization study is listed in Table 2. They are also an excellent, low-cost source for enzyme production [97].
An enzymatic approach can overcome the environmental impact of traditional processes and make the processes sustainable and cost-effective. In a very recent comprehensive review, it is reported that significant developments can be expected for enzyme applications in the fish and seafood industries [98] in the near future. Hydrolysates improve the immune system with differential influences on the immune function. Interesting study for several applications in the health food, pharmaceutical, and nutraceutical industries

Fine Chemistry and Lab Techniques
Biocatalytic procedures using marine enzymes for the production of fine chemicals are an important aspect of this review. The production and manipulation of complex biomolecules benefited from important biotechnological features characterizing biocatalyzed reactions with respect to the use of purely chemical-based methods [99].
Technological improvements (metagenomics) applied to bioprospecting in understudied environments help to identify a greater repertoire of novel biocatalysts with complementarity about properties (stereochemistry, resistance, etc.). Marine enzymes offer hyperthermostability, salt tolerance, barophilicity, cold adaptability, chemoselectivity, regioselectivity and stereoselectivity [4], thus acting as useful and new alternatives to terrestrial biocatalysts in use. Particular importance is represented by enzymes showing resistance to organic solvents, with the examples from marine environments mostly related to halophilic proteins (salt reduces water activity, like organic solvent systems), as analyzed in a recent, comprehensive review article [100].
As for carbohydrate-active enzymes, selective and easy manipulation of structurally complex marine polysaccharides provides homogeneous core blocks (oligosaccharides) for analysis and hemisynthesis. This constitutes the core of a sustainable process when using renewable resources.
Other preminent examples of enzymatic treatments as a tool to improve the extraction efficiency of specific bioactive compounds from seaweeds were recently reviewed [101] and laboratory techniques in the preparation of compounds for further research are discussed in a recent report [102] listing enzymes for the functionalization of chitosan such as polyphenoloxidases (PPO) (tyrosinases, laccases) and peroxidases (POD); examples from the marine environment are indicated.
Due to our specific design of search terms in querying literature databases, the coverage of articles dealing with the simple prospecting, isolation and identification of new marine biocatalyst(s) is partial; however, however those found are included in this section. Indeed, most articles listed in Table 3 [103-134] focused on bioprocesses for the synthesis of useful products and on enzymatic routes adopted for setting up laboratory techniques to study marine complex biomolecules (e.g., improving extraction, digestion of polysaccharides to simple components for structural determination, etc.). Various examples are found in the literature of the synthesis and hydrolysis of glycosidic bonds. Entry 8, Table 3 [110] is only one of the examples of synthetic strategies for the production of interesting products (enzymatically glycosylated natural lipophilic antioxidants). Polymers with synthetic carbohydrates have a wide range of applications in medical biotechnology as new biomaterials [135] and carbohydrate-active hydrolases can also be applied; moreover, these biocatalysts are important for the synthesis of a number of novel dietary carbohydrates in food technology, for the production of chromophoric oligosaccharides of strictly defined structure as valuable biochemical tools, etc. Other numerous applications oriented to vegetal waste treatment in recovering useful materials are reported in the section devoted to biorefinery.
Tabulation of primary articles in this section (Table 3) is based on biocatalyst(s) used, product(s) obtained with the biocatalyzed process, or evaluating of marine feedstock(s); comments about the contents of the article are also reported in the notes in Table 3. Entries 1-18 (Table 3) are related to carbohydrate-active hydrolases, while a few (entries 19-23) are listed for ester hydrolysis and proteolytic activities (entries 31 and 32).   Little is known about the distribution and diversity of Candida genus in marine environments, with only a few species isolated from these environments [136]. C. rugosa and C. antarctica are among the terrestrial yeasts, so biocatalysis-related articles dealing with these quite famous commercial lipases are not listed here. Interesting examples of oxidoreductases are also found in the literature, both for direct biocatalytic applications (including examples of immobilized enzymes) and for biogenetically related studies (see entries 24-30, Table 3).
Finally, it is worth mentioning in this section a remarkable review focused on the biosynthesis of oxylipins in non-mammals. Biocatalysts involved in pathways related to these biomolecules carry interesting and unusual catalytic properties for biocatalysis. A detailed biocatalytic knowledge of enzymatic catalysis in these reactions is needed to plan a possible direct in vitro biocatalytic lab-scale production of useful products [137]. With the similar aim of increasing knowledge about natural enzymes with interesting features, a review was compiled on the enzymatic breakage of dimethylsulfoniopropionate [138]. The compound, a zwitterionic osmolyte produced by corals, marine algae and some plants in massive amounts (ca. 10 9 tons per year), is transformed by marine microbes and bioprocesses involving this molecule and derivatives are of interest for assessing the ability of relevant enzymes to realize these transformations.

Sediments and Bioremediation
Among the studies on marine sources for enzymes, the fields of marine biomarkers and bioremediation applications are of high significance in this context [139].
In the list of parameters that are usually considered when assessing exposure to environmental pollutants in aquatic ecosystems, biotransformation enzymes (phase I and II), biotransformation products and stress proteins are of high interest for enzymatic processing. It is of value, for example, that a unique set of protein expression (signature) for exposure to different chemical compounds has been recognized for Mytilus edulis and the expressed proteins identified participate in αand β-oxidation pathways, xenobiotic and amino acid metabolism, cell signalling and oxyradical metabolism [140]. Bioremediation as an enabling technology exploits naturally occurring organisms that with their metabolic ability are able to transform toxic substances in less hazardous compounds that in turn are included in biogeochemical cycles. Stereochemical aspects play an important role due to the fact that homochirality appears to be a requirement for the functioning of enzymes with specific (partial) incorporation of stereoforms. Enantioselective chromatographic separation of chiral environmental xenobiotics is covered in an interesting review. The study includes microbial transformation of chiral pollutants in aquatic ecosystems, enzymatic transformations, etc. [141]. An additional report states the effectiveness of enzymatic processes in bioremediation even though limited application is evidenced with respect to stability and the cost of biocatalysts. Marine enzymes are seen as a solution to this challenge. In particular, marine fungi and their laccases are used in the textile industry, mostly to deal with salty effluents [142].
A recent report of a European project in this field characterizes novel hydrocarbon degrading microbes isolated from the southern side of the Mediterranean Sea. Exploiting and managing the diversity and ecology of microorganisms thriving in these polluted sites is a major objective in terms of increasing knowledge of the bioremediation potential of these poorly investigated sites [143].
In Table 4 primary articles in the field are selected , indicating the biocatalyst(s) or organism(s) exploited and details of application, with notes about the results and importance of the work.  Catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase, glutathione S-transferases in wild populations of mussels (Mytilus galloprovincialis)

Study of biochemical response to to petrochemical environmental contamination
Environmental monitoring programmes to get data that could be used as a baseline reference during oil accidents 18/2007 [161] Acetylcholinesterase, catalase, and glutathione-S-transferase (GST) of blue mussels (Mytilus edulis) Study of specific reaction to exposure to nodularin Acetylcholinesterase activity, catalase (CAT) activity and glutathione-S-transferase (GST) in blue mussels (Mytilus edulis) exposed to an extract made of natural cyanobacterial mixture containing toxic cyanobacterium Nodularia spumigena

19/2007 [162] Induction of biotransformation enzymes in Sparus aurata
Relationship between specific molecular processes (induction of enzymes) and the behavioral performance of fish is of great interest in understanding the impact of PAHs at increasing levels of biological complexity The study investigates biochemical response to phenantrene in Sparus aurata

Others
Searching within records used in this review for the names of class of enzymes, oxidoreductases, transferases, hydrolases and lyases are equally represented, while isomerases and ligases are less often used as keywords. However, only 30% of the total records contain such names for classes, with common names of biocatalysts more often adopted in titles, abstracts and keywords. Thus, in this section a few other topics, difficult to insert in previous sections, are covered.
Enzymes acting on 1,4 glycosidic bonds between galacturonic acid residues in pectin were investigated for the improvement of banana fiber processing. At least one actinomycete strain of Streptomyces lydicus collected from estuarine and marine areas in India was found to be a potent producer of polygalacturonase [169].
In a study to reduce the cost of aquaculture of fish cobia by adding crustacean processing waste, an investigation of the endogenous chitinolytic enzymes in cobia was conducted. It suggested substantial endogenous production of enzymes of the chitinolytic system and that the activity from chitinolytic bacteria was not significant [170].
An interesting study based on enzymology tools was conducted for demonstrating the enhanced mixing processes between the sediment and the overlying waters of the Delaware Estuary. The authors used fluorescently labeled polysaccharides to determine the effects of suspended sediment transport on water column hydrolytic activities [171].
In another report, a rapid and easy-to-use set, composed of semi-quantitative kits, was adopted for the investigation of the heterotrophic bacterial community in meadows of Posidonia oceanica during environmental surveys. Although the set is composed of known kits (ApiZym galleries, Biolog microplates and BART™ tests) principal enzymatic activities, metabolic capabilities and benthic mineralisation processes were all studied [172].
A particular aspect is related to carbonic anhydrases; it is worth mentioning in this review since it was recently discussed in a book devoted to these enzymes from extremophiles and the possible biotechnological applications for which they can be used [173]. Among the CO 2 sequestration methods proposed in order to capture and concentrate CO 2 from combustion gases, the biomimetic approach [174] could indeed benefit from the diversity of marine carbonic anhydrases. However, these metallo-enzymes are also discussed for their potential as novel biomarkers in environmental monitoring and the development of biosensors for metals [175], and another interesting aspect, encompassing at least two of the fields listed in this review, is the investigation of other enzymatic activities possessed by carbonic anhydrases [176].

Conclusions
One of the recent transnational calls of ERA-NET (ERA-MBT), an action funded under the EU FP7 program, was focused on biorefinery and entitled "The development of biorefinery processes for marine biomaterials". The projects were required to develop the production of a large number of different products and novel processes through the application of biotechnological knowledge. Further technological developments to improve the integration and optimisation of the processing steps were required. In this realm seaweed biomass seems to have great potential as a raw material in a biobased economy, offering advantages such as no competion with food production, absence of fertilisers or pesticides in the value-chain and positive relapses removing an excess of nutrients from marine environments. However, a few key areas where development is still needed are technology for the improvement of large-scale cultivation and fractionation, and the identification of new marine microbial strains to break down macroalgal polysaccharide.
The use of macroalgae in food preparation is a classical topic in food applications found when inspecting the primary articles listed in Table 2. Macroalgae contain a high concentration of minerals, vitamins, trace elements and fibre and have low fat content. Different projects studying these aspects are in development. It is of interest that a close inspection of the affiliations of the corresponding authors of all articles in Table 2 reveals that Spain and France are the top two countries where research efforts were published; however, many countries produce interesting research results.
As for fine chemistry and laboratory techniques, different case studies recently illustrated the importance of biocatalysis, considering the specificities of marine enzymes with respect to their terrestrial counterparts [177]. Ketone reduction and epoxide hydrolases useful in organic synthesis appeared to be central for stereochemical aspects. Access to bioactive aldehydes with lipooxygenases and lipases actions and biodegradation of marine pollutants are covered, along with other lipolytic activities used for enantioselective hydrolysis. From the overall analysis and examples reported, the strategy regarding the potential of marine habitats is clear. It is important to report the note of two editors in a special issue dedicated to biocatalysis in Current Opinion in Chemical Biology; they stated that "At least a third of this planet's biomass resides in the oceans, and the rules of the marine biochemical game seem to be fundamentally different than those described in our biochemistry textbooks." [178].
In the bioremediation field, [141] is very stimulating, pointing out the importance of stereoselectivity in aquatic ecosystem biotransformations and reporting on the great impact that the use of cyclodextrins in chiral gas-chromatography had in widening this knowledge. All the information reported by the studies on processes is of great interest to biocatalysis practitioners, starting from pioneering investigations into the distribution of atropisomeric PCBs in the marine environment. Marine bacteria specialising in the degradation of hydrocarbons have been isolated from polluted seawater and some of these bacteria can grow on these substrates; they represent an extraordinary archive of mono-and dioxygenases, oxidases, dehydrogenases and other enzymatic activities that can be applied in regio-and stereoselective biocatalysis. There is a gap between the general knowledge from the studies in this section and the specificity/suitability required for preparative enzymatic processes; bridging this gap could shed more light on the useful features of the enzymes involved in this type of pollutant biotransformation, thus enabling more effective application.
Fewer than half of the results from our literature search are cited in the references list. About 20% of the hits are represented by books or reference works that were hardly used here. Additionally, only four scientific journals hosted more than 10 articles of the remaining corpus: Bioresource Technology, Food Chemistry, Process Biochemistry and Chemosphere together account fornot far off 100 articles. The top specialist marine-oriented journal was Algal Research, with only eight papers. Inserting representative primary articles in the tables and excluding the ones that do not belong resulted in the current total number of references. Therefore, scientific interest in marine enzymatic processing can be considered successfully published in non-specialized journals such as the ones cited above; the separation of fields, as adopted here, is only for ease of discussion. This has already been mentioned above for marine biomass-centered studies that could have been listed under food applications or biorefinery, or for enzymatic activities used for polysaccharide manipulation that could have been listed under fine chemistry and lab techniques instead of biorefinery.
In conclusion, all these aspects point to a final consideration of the importance of an interdisciplinary network in setting up successful research projects enabling the identification of an arsenal of enzymes and pathways greatly in demand for biotechnological applications. In continuing this research effort, further refining of the scientific literature could be of interest; exploration of the fields individuated above should be continued in depth, in specialized journals, in a manner that could help to reveal sub-fields along with more details pointing to a single process with room to discuss a single enzymatic activity.